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  • ‹ Back to Cyan and green fluorescent proteins
  • AcGFP1 fluorescent protein
  • ZsGreen1 fluorescent protein
  • GFP & GFPuv fluorescent proteins
  • AmCyan1 fluorescent protein
Selection guides Fluorescent protein quick guide
Antibodies to GFP variants, mCherry and other fluorescent proteins Fluorescent protein antibodies
Home › Products › Gene function › Fluorescent proteins › Fluorescent protein plasmids › Cyan and green fluorescent proteins › ZsGreen1 fluorescent protein

Fluorescent protein plasmids

  • Cyan and green fluorescent proteins
    • AcGFP1 fluorescent protein
    • ZsGreen1 fluorescent protein
    • GFP & GFPuv fluorescent proteins
    • AmCyan1 fluorescent protein
  • Red fluorescent proteins
    • mCherry fluorescent protein
    • DsRed-Monomer fluorescent protein
    • DsRed2 fluorescent protein
    • DsRed-Express and DsRed-Express2 fluorescent proteins
    • tdTomato fluorescent protein
    • AsRed2 fluorescent protein
    • mStrawberry fluorescent protein
  • Far-red fluorescent proteins
    • E2-Crimson fluorescent protein
    • HcRed1 fluorescent protein
    • mRaspberry fluorescent protein
    • mPlum fluorescent protein
  • Orange and yellow fluorescent proteins
    • mOrange2 fluorescent protein
    • mBanana fluorescent protein
    • ZsYellow1 fluorescent protein
  • Photoactivatable and photoswitchable proteins
    • Dendra2 fluorescent protein
    • Timer fluorescent protein
    • PAmCherry fluorescent protein
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Selection guides Fluorescent protein quick guide
Antibodies to GFP variants, mCherry and other fluorescent proteins Fluorescent protein antibodies

ZsGreen1 fluorescent protein

Living Colors ZsGreen1 is an exceptionally bright green fluorescent protein derived from a Zoanthus sp. reef coral (Matz et al. 1999) that has been modified for high solubility, bright emission, and rapid chromophore maturation. ZsGreen1 is the brightest commercially available green fluorescent protein—up to 4X brighter than EGFP—and is ideally suited for whole-cell labelling, promoter-reporter studies, or as a transfection control.

Living Colors ZsGreen1 is an exceptionally bright green fluorescent protein derived from a Zoanthus sp. reef coral (Matz et al. 1999) that has been modified for high solubility, bright emission, and rapid chromophore maturation. ZsGreen1 is the brightest commercially available green fluorescent protein—up to 4X brighter than EGFP—and is ideally suited for whole-cell labelling, promoter-reporter studies, or as a transfection control. The protein naturally forms a tetramer and is therefore not optimal for development of fusion proteins; we recommend AcGFP1, a true monomer, for such applications.

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Cat. # Product Size Price License Quantity Details
632598 Living Colors® ZsGreen Monoclonal Antibody 100 uL USD $506.00

The Living Colors ZsGreen Monoclonal Antibody was raised in mouse against recombinant full-length Zoanthus sp. green fluorescent protein (ZsGreen), and Protein A-purified. In Western blot applications, it specifically recognizes ZsGreen and does not cross-react with AcGFP1, AmCyan, ZsYellow, DsRed-Express, mCherry, tdTomato, Dendra2, or DsRed2.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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632598: Living Colors ZsGreen Monoclonal Antibody

632598: Living Colors ZsGreen Monoclonal Antibody
632566 pLVX-ZsGreen1-C1 Vector 10 ug USD $705.00

This lentiviral expression vector encodes the ZsGreen1 fluorescent protein tag. The coding sequence for this very bright green fluorescent protein (2.5x brighter than EGFP) was modified to include human codon usage preferences, which allow improved expression in mammalian cells. Inserting a cDNA in the MCS downstream of the ZsGreen1 coding sequence joins your protein of interest to the C-terminus of the tag, and allows the fusion protein to be tracked and studied in transduced cells. To package the vector into high-titer, replication-incompetent lentivirus, we recommend using Lenti-X Packaging Single Shots and the Lenti-X 293T Cell Line. The resulting lentivirus can then be used to transduce virtually any mammalian cell type.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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Lentiviral vectors with fluorescent proteins

Lentiviral vectors with fluorescent proteins

Lentiviral vectors with fluorescent proteins. Lenti-X vectors contain sequence elements that facilitate lentiviral packaging and/or boost transgene expression, including the LTRs, packaging signal (Ψ), Rev response element (RRE), and central polypurine tract/central termination sequence (cPPT/CTS) from HIV-1; and the woodchuck hepatitis virus post-transcriptional regulatory element (WPRE). Vectors can express your protein fused at its N- or C- terminus to either a green (AcGFP1) or red (DsRed-Monomer) fluorescent protein tag, or coexpress it as a separate protein along with ZsGreen1 (shown), mCherry, or tdTomato.

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632566: pLVX-ZsGreen1-C1 Vector

632566: pLVX-ZsGreen1-C1 Vector
632565 pLVX-ZsGreen1-N1 Vector 10 ug USD $705.00

This lentiviral expression vector encodes the ZsGreen1 fluorescent protein tag. The coding sequence for this very bright green fluorescent protein (2.5x brighter than EGFP) was modified to include human codon usage preferences, which allow improved expression in mammalian cells. Inserting a cDNA in the MCS upstream of the ZsGreen1 coding sequence joins your protein of interest to the N-terminus of the tag, and allows the fusion protein to be tracked and studied in transduced cells.

To package the vector into high-titer, replication-incompetent lentivirus, we recommend using Lenti-X Packaging Single Shots and the Lenti-X 293T Cell Line. The resulting lentivirus can then be used to transduce virtually any mammalian cell type.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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632565: pLVX-ZsGreen1-N1 Vector

632565: pLVX-ZsGreen1-N1 Vector

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Lentiviral vectors with fluorescent proteins

Lentiviral vectors with fluorescent proteins

Lentiviral vectors with fluorescent proteins. Lenti-X vectors contain sequence elements that facilitate lentiviral packaging and/or boost transgene expression, including the LTRs, packaging signal (Ψ), Rev response element (RRE), and central polypurine tract/central termination sequence (cPPT/CTS) from HIV-1; and the woodchuck hepatitis virus post-transcriptional regulatory element (WPRE). Vectors can express your protein fused at its N- or C- terminus to either a green (AcGFP1) or red (DsRed-Monomer) fluorescent protein tag, or coexpress it as a separate protein along with ZsGreen1 (shown), mCherry, or tdTomato.

632541 pmR-ZsGreen1 Vector 20 ug USD $596.00

pmR-ZsGreen1 is a mammalian expression vector designed to constitutively coexpress a microRNA (miRNA) of interest and the ZsGreen1 fluorescent protein. ZsGreen1 is a human codon-optimized variant of the Zoanthus sp. green fluorescent protein, ZsGreen, which has been engineered for brighter fluorescence and higher expression in mammalian cells. The unmodified vector will express ZsGreen1 in mammalian cells.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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The pmR-mCherry and pmR-ZsGreen1 vectors will coexpress a fluorescent protein and a miRNA sequence that is embedded in the 3' UTR of the vector’s mRNA transcript

The pmR-mCherry and pmR-ZsGreen1 vectors will coexpress a fluorescent protein and a miRNA sequence that is embedded in the 3' UTR of the vector’s mRNA transcript

The pmR-mCherry and pmR-ZsGreen1 vectors will coexpress a fluorescent protein and a miRNA sequence that is embedded in the 3' UTR of the vector’s mRNA transcript. miRNA expression can be selected for and/or verified in transfected cells by monitoring red or green fluorescence.

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The pmR-mCherry and pmR-ZsGreen1 Vectors

The pmR-mCherry and pmR-ZsGreen1 Vectors

The pmR-mCherry and pmR-ZsGreen1 Vectors. Map of the vectors (Panel A). Cells transfected with the vectors express your miRNA and exhibit red or green fluorescence (Panel B).

632448 pZsGreen1-N1 Vector 20 ug USD $615.00

pZsGreen1-N1 is a human codon-optimized expression vector that encodes a variant of the Zoanthus sp. green fluorescent protein, ZsGreen1, which has been engineered for brighter fluorescence and higher expression in mammalian cells. pZsGreen1-N1 allows genes cloned into the multiple cloning site (MCS) upstream of the ZsGreen1 coding sequence to be expressed as fusions to the N-terminus of the fluorescent protein. The unmodified vector will express ZsGreen1 in mammalian cells.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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Fluorescent whole-body image of a mouse with bilateral solid tumors derived from estrogen receptor-positive MCF7 human breast cancer cells

Fluorescent whole-body image of a mouse with bilateral solid tumors derived from estrogen receptor-positive MCF7 human breast cancer cells

Fluorescent whole-body image of a mouse with bilateral solid tumors derived from estrogen receptor-positive MCF7 human breast cancer cells. Note: the presence of ZsGreen-positive cells in tumors, lymphatic vessels (LV), and inguinal and axillary lymph nodes (LN).

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632448: pZsGreen1-N1 Vector

632448: pZsGreen1-N1 Vector

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Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2, and HcRed1

Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2, and HcRed1

Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2,  and HcRed1.

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Schematic for fluorescent fusion protein vectors

Schematic for fluorescent fusion protein vectors

Schematic for fluorescent fusion protein vectors. Panel A. N-terminal fluorescent protein (FP) vector. Panel B. C-terminal fluorescent protein vector.

632447 pZsGreen1-C1 Vector 20 ug USD $615.00

pZsGreen1-C1 is a human codon-optimized expression vector that encodes a variant of the Zoanthus sp. green fluorescent protein, ZsGreen1, which has been engineered for brighter fluorescence and higher expression in mammalian cells. pZsGreen1-C1 allows genes cloned into the multiple cloning site (MCS) downstream of the ZsGreen1 coding sequence to be expressed as fusions to the C-terminus of the fluorescent protein. The unmodified vector will express ZsGreen1 in mammalian cells.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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632447: pZsGreen1-C1 Vector

632447: pZsGreen1-C1 Vector

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Fluorescent whole-body image of a mouse with bilateral solid tumors derived from estrogen receptor-positive MCF7 human breast cancer cells

Fluorescent whole-body image of a mouse with bilateral solid tumors derived from estrogen receptor-positive MCF7 human breast cancer cells

Fluorescent whole-body image of a mouse with bilateral solid tumors derived from estrogen receptor-positive MCF7 human breast cancer cells. Note: the presence of ZsGreen-positive cells in tumors, lymphatic vessels (LV), and inguinal and axillary lymph nodes (LN).

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Schematic for fluorescent fusion protein vectors

Schematic for fluorescent fusion protein vectors

Schematic for fluorescent fusion protein vectors. Panel A. N-terminal fluorescent protein (FP) vector. Panel B. C-terminal fluorescent protein vector.

632446 pZsGreen Vector 20 ug USD $615.00

pZsGreen is a prokaryotic expression vector, which encodes Zoanthus sp. green fluorescent protein (ZsGreen). The ZsGreen coding sequence carried in pZsGreen retains the wild-type codon usage. In this vector, the ZsGreen gene is flanked by separate and distinct MCS regions at the 5' and 3' ends so that the ZsGreen gene can be subcloned into other expression vectors. This vector is primarily intended to serve as a convenient source of ZsGreen cDNA.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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Four-color visualization of Living Colors Fluorescent Proteins

Four-color visualization of Living Colors Fluorescent Proteins
Four-color visualization of Living Colors Fluorescent Proteins. HEK 293 clonal cell lines stably expressing AmCyan1, ZsGreen1, ZsYellow1, or HcRed1 were mixed and plated in the same culture dish and imaged at 20X magnification.

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632446: pZsGreen Vector

632446: pZsGreen Vector

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Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2, and HcRed1

Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2, and HcRed1

Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2,  and HcRed1.

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Basic fluorescent protein vector map

Basic fluorescent protein vector map

Basic fluorescent protein vector map. Use this bacterial expression vector as a source of the fluorescent protein gene. Note: There is a stop codon at the 5' end of the 3' MCS. The 3' MCS should not be used for cloning.

632520 pRetroX-IRES-ZsGreen1 Vector 20 ug USD $593.00

The pRetroX-IRES-ZsGreen1 vector is a bicistronic retroviral expression vector containing ZsGreen1 as a marker for transfection/infection efficiency. A gene of interest is inserted into the MCS located upstream of the encephalomyocarditis virus (EMCV) internal ribosomal entry site (IRES). The IRES sequence allows the protein of interest and ZsGreen1 to be independently translated from the same mRNA transcript. ZsGreen1 is a human codon-optimized variant of the Zoanthus sp. green fluorescent protein (ZsGreen) that has been engineered for brighter fluorescence and higher expression in mammalian cells.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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632520: pRetroX-IRES-ZsGreen1 Vector

632520: pRetroX-IRES-ZsGreen1 Vector

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Mechanism of IRES bicistronic expression from Retro-X retroviral vectors

Mechanism of IRES bicistronic expression from Retro-X retroviral vectors

Mechanism of IRES bicistronic expression from Retro-X retroviral vectors. The internal ribosome entry site (IRES) permits a protein of interest and a fluorescent protein to be independently translated from the same mRNA. SD = Splice Donor. SA = Splice Acceptor. ψ = packaging signal. LTR = long terminal repeat.

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Retro-X retroviral vector maps

Retro-X retroviral vector maps

Retro-X retroviral vector maps. IRES-containing bicistronic retroviral vectors allow you to express two target genes (Vector A: pQCXIX) or a target gene and an antibiotic resistance gene (Vector B: pQCXIH,pQCXIN and pQCXIP). Fluorescent-fusion retroviral vectors express your protein of interest fused to a fluorescent protein (Vector C: pRetroQ-AcGFP1 N1/C1 and pRetroQ-DsRed Monomer N1/C1). Knockout RNAi-Ready pSIREN-RetroQ retroviral vectors are prelinearized and ready to accept a dsDNA oligonucleotide encoding a shRNA of your choice, (Vectors D and E: RNAi-Ready pSIREN-RetroQ-DsRedExpress or pSIREN-RetroQ-ZsGreen and RNAi-Ready pSIREN-RetroQ). To express shRNA under tight, inducible control, use the knockout Tet series of retroviral vectors (Vector F: Knockout RNAi-Ready pSIREN-RetroQ-TetH and pSIREN-RetroQ TetP).

632478 plRES2-ZsGreen1 Vector 20 ug USD $615.00

pIRES2-ZsGreen1 is a bicistronic expression vector containing ZsGreen1 as a marker for transfection efficiency. It encodes a human-codon-optimized variant of the Zoanthus sp. green fluorescent protein, ZsGreen, which has been engineered for brighter fluorescence and higher expression in mammalian cells. A gene of interest is inserted into the MCS located upstream of the encephalomyocarditis virus (EMCV) internal ribosomal entry site (IRES). The IRES sequence allows the gene of interest and ZsGreen1 to be translated simultaneously from the same mRNA transcript.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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Map of the fluorescent pIRES2 bicistronic expression vectors

Map of the fluorescent pIRES2 bicistronic expression vectors
Map of the fluorescent pIRES2 bicistronic expression vectors.

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Expression of two proteins from a single mRNA transcript

Expression of two proteins from a single mRNA transcript
Expression of two proteins from a single mRNA transcript. A fluorescent protein is translated from an internal ribosome entry site (IRES).

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632478: plRES2-ZsGreen1 Vector

632478: plRES2-ZsGreen1 Vector

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Use of AcGFP1 for fusions and fluorescence microscopy applications

Use of AcGFP1 for fusions and fluorescence microscopy applications

Use of AcGFP1 for fusions and fluorescence microscopy applications. Panels A and B. Activation of Protein Kinase C alpha was monitored with Living Colors AcGFP1. Panel A. HEK 293 cells were stably transfected with a plasmid encoding AcGFP1 fused to PKC alpha. Panel B. Cells were induced with 1.5 µg/ml PMA for 3 min. The PKC alpha-AcGFP1 fusion moves from the cytosol to the plasma membrane, a result consistent with the known mobilization pattern of PKC alpha. Panel C. HeLa cells were transiently transfected with pAcGFP1-Actin and visualized by fluorescence microscopy.

632187 pLVX-IRES-ZsGreen1 Vector 10 ug USD $705.00

The pLVX-IRES-ZsGreen1 Vector is a bicistronic lentiviral expression vector that can be used to generate high-titer lentivirus for transducing dividing or non-dividing mammalian cells. The vector contains an internal ribosomal entry site (IRES) which allows a gene-of-interest and the ZsGreen1 fluorescent protein to be simultaneously coexpressed from a single mRNA transcript. When used with Lenti-X Packaging Single Shots (e.g. Cat. No. 631275) and our Lenti-X 293T Cell Line (Cat. No. 632180), the vector generates high titers of replication-incompetent, VSV-G pseudotyped lentivirus.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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Lentiviral vectors with fluorescent proteins

Lentiviral vectors with fluorescent proteins

Lentiviral vectors with fluorescent proteins. Lenti-X vectors contain sequence elements that facilitate lentiviral packaging and/or boost transgene expression, including the LTRs, packaging signal (Ψ), Rev response element (RRE), and central polypurine tract/central termination sequence (cPPT/CTS) from HIV-1; and the woodchuck hepatitis virus post-transcriptional regulatory element (WPRE). Vectors can express your protein fused at its N- or C- terminus to either a green (AcGFP1) or red (DsRed-Monomer) fluorescent protein tag, or coexpress it as a separate protein along with ZsGreen1 (shown), mCherry, or tdTomato.

Required Products

Cat. # Product Size Price License Quantity Details
631275 Lenti-X™ Packaging Single Shots (VSV-G) 16 Rxns USD $1071.00

License Statement

ID Number  
63 Use of this product is covered by one or more of the following U.S. Patent Nos. and corresponding patent claims outside the U.S.: 8,562,966, 8,557,231. This product is intended for research purposes only. It may not be used for (i) any human or veterinary use, including without limitation therapeutic and prophylactic use, (ii) any clinical use, including without limitation diagnostic use, (iii) screening of chemical and/or biological compounds for the identification of pharmaceutically active agents (including but not limited to screening of small molecules), target validation, preclinical testing services, or drug development. Any use of this product for any of the above mentioned purposes requires a license from the Massachusetts Institute of Technology.
259 This Product is protected by one or more patents from the family consisting of: JP6454352 and any corresponding patents, divisionals, continuations, patent applications and foreign filings sharing common priority with the same family.

Lenti-X Packaging Single Shots (VSV-G) provide an extremely simple and consistent one-step method for producing high-titer lentivirus. No additional transfection reagent is needed because Lenti-X Packaging Single Shots (VSV-G) consist of pre-aliquoted, lyophilized, single tubes of Xfect Transfection Reagent premixed with an optimized formulation of VSV-G pseudotyped Lenti-X lentiviral packaging plasmids. High-titer virus is produced by simply reconstituting this mixture with your lentiviral vector of choice in sterile water and adding it to 293T cells, e.g., Lenti-X 293T Cells (Cat. # 632180), in a 10 cm dish.

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The Lenti-X Packaging Single Shots (VSV-G) protocol

The Lenti-X Packaging Single Shots (VSV-G) protocol
The Lenti-X Packaging Single Shots (VSV-G) protocol.

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Consistent, high-efficiency transfections lead to high titers

Consistent, high-efficiency transfections lead to high titers

Consistent, high-efficiency transfections lead to high titers. A lentiviral vector containing the ZsGreen1 gene was packaged according to the Lenti-X single shots protocol in four independent experiments. Briefly, 7 µg of pLVX-ZsGreen1 plasmid was added to each of four Lenti-X single shots; the tubes were vortexed for 20 sec and incubated at room temperature for 10 min. Then, the mixture was added to cultured Lenti-X 293T cells that were approximately 80% confluent. 48 hours after transfection, the cells were imaged by fluorescence microscopy (Panel A, top) and light microscopy (Panel A, bottom). After images were taken, the supernatant was harvested and used infect HT1080 cells for titer determination (Panel B, IFU/ml).

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High-titer virus was produced regardless of the lentiviral vector backbone with Lenti-X packaging single shots

High-titer virus was produced regardless of the lentiviral vector backbone with Lenti-X packaging single shots

High-titer virus was produced regardless of the lentiviral vector backbone with Lenti-X packaging single shots. A CMV ZsGreen1 expression cassette was cloned into several lentiviral vector backbones. These vectors were then packaged into lentivirus using the Lenti-X packaging single shots following the provided protocol. Briefly, 7 µg of pLVX-ZsGreen1 plasmid was added to each of four Lenti-X single shots, and the tubes were vortexed for 20 sec and incubated at room temperature for 10 min. Then, the mixture was added to cultured Lenti-X 293T cells that were approximately 80% confluent. After 48 hours, titer was determined using several methods. To determine infectivity, the supernatant was harvested and used to infect HT1080 cells (Flow Cytometry). Harvested viral supernatants were also analyzed by RT-PCR to quantify viral genome copies (qRT-PCR, Lenti-X qRT-PCR Titration Kit), ELISA to measure p24 (p24 ELISA, Lenti-X p24 Rapid Titer Kit), and by a rapid lentiviral detection method (Lenti-X GoStix).

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A comparison of fourth- and third-generation lentiviral packaging systems

A comparison of fourth- and third-generation lentiviral packaging systems

A comparison of fourth- and third-generation lentiviral packaging systems . Our Lenti-X Packaging single shots utilize a packaging system that consists of five separate components (Panel A), mixed in proprietary proportions for optimized packaging activity. The separation of the gag, pol, and env genes effectively reduces the incidence of RCL (Wu et al., 2000). High levels of expression of essential viral components are driven by the Tet-Off and Tat transactivators, which induce a cascade of expression that results in high titers of lentivirus. The pol gene is fused to vpr to ensure transport of the reverse transcriptase/integrase protein into the recombinant lentiviral particle. Not all vector elements are shown. Other 3rd generation lentiviral packaging systems (Panel B) generate lower titers, do not contain separate gag and pol sequences, and do not use a transactivation cascade mechanism.

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631275: Lenti-X Packaging Single Shots (VSV-G)

631275: Lenti-X Packaging Single Shots (VSV-G)
632180 Lenti-X™ 293T Cell Line 1 mL USD $422.00

License Statement

ID Number  
406 This product is the subject of a technology license agreement for internal research use only. Use of this product other than for research use may require additional licenses. Information on license restrictions or for uses other than research may be obtained by contacting licensing@takarabio.com.

The Lenti-X 293T Cell Line is a subclone of the transformed human embryonic kidney cell line, HEK 293, which is highly transfectable and supports high levels of viral protein expression. When transfected with Lenti-X Packaging Single Shots and a lentiviral vector, these cells are capable of producing lentiviral titers as high as >108 ifu/ml, as determined by flow cytometry. The cell line also constitutively expresses the simian virus 40 (SV40) large T antigen.

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632180: Lenti-X 293T Cell Line

632180: Lenti-X 293T Cell Line

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Clontech's lentiviral packaging system (Panel A) and a lentiviral packaging system from a leading competitor (Panel B) were each used to generate viral supernatants from their respective lentiviral system vectors that were engineered to express the ZsGreen1 fluorescent protein

Clontech's lentiviral packaging system (Panel A) and a lentiviral packaging system from a leading competitor (Panel B) were each used to generate viral supernatants from their respective lentiviral system vectors that were engineered to express the ZsGreen1 fluorescent protein
Clontech's lentiviral packaging system (Panel A) and a lentiviral packaging system from a leading competitor (Panel B) were each used to generate viral supernatants from their respective lentiviral system vectors that were engineered to express the ZsGreen1 fluorescent protein. As little as 10 µl of supernatant from Lenti-X transduced the majority of these HeLa cells, whereas 10 µl of supernatant from the other system transduced only a small percentage of the cells. Transduced cells were quantified by flow cytometry.

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Transduction of neural progenitor cells by Lenti-X lentivirus

Transduction of neural progenitor cells by Lenti-X lentivirus

Transduction of neural progenitor cells by Lenti-X lentivirus. Recombinant lentivirus for expressing ZsGreen1 was produced using Lenti-X virus and used to transduce normal human neural progenitor cells. A single transduced cell is shown under phase contrast microscopy (Panel A) and fluorescence microscopy (Panel B).

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High-titer lentivirus production

High-titer lentivirus production

High-titer lentivirus production. Lenti-X 293T cells were transduced with the indicated volumes (µl) of lentiviral packaging supernatant generated with the Lenti-X Expression System and then selected with puromycin for 9 days to allow the formation of the resistant colonies, which were then stained with crystal violet.

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293T cell line for higher titers

293T cell line for higher titers

293T cell line for higher titers. We used our fourth-generation lentiviral packaging system and one of our pLVX-lentiviral vectors to compare the virus production of the Lenti-X 293T Cell Line to that of two other commonly used HEK 293-based cell lines. Lenti-X 293T cells clearly outperformed the other cell lines—producing over 6X more virus than 293FT cells and up to 30X more virus than the parental HEK 293 cell line.

631982 pLVX-EF1alpha-IRES-ZsGreen1 Vector 10 ug USD $705.00

pLVX-EF1α-IRES-ZsGreen1 is a bicistronic lentiviral expression vector that can be used to generate high-titer lentivirus for transducing virtually any dividing or nondividing mammalian cell type, including primary and stem cells. The vector contains an internal ribosomal entry site (IRES) that allows a gene-of-interest and the ZsGreen1 fluorescent protein to be simultaneously coexpressed from a single mRNA transcript. Expression of the transcript is driven by the constitutively active human elongation factor 1 alpha (EF1α) promoter.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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631982: pLVX-EF1alpha-IRES-ZsGreen1 Vector

631982: pLVX-EF1alpha-IRES-ZsGreen1 Vector

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Lentiviral vector systems with EF-1 alpha promoters

Lentiviral vector systems with EF-1 alpha promoters
Lentiviral vector systems with EF-1 alpha promoters. Achieve robust, constitutive, long-term expression of your gene of interest in cell types in which CMV promoters are often silenced, such as hematopoietic and stem cells.

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Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter. The mouse embryonic stem cell lines E14 (Panel A) and D3 (Panel B) were transduced by Lenti-X lentivirus, expressing AcGFP1 either under the control of the CMV promoter or the Elongation factor alpha (EF-1 alpha) promoter. The expression level of AcGFP1 in infected cells five days postinfection was monitored by FACS analysis using the FL1 channel. The expression of AcGFP1 driven by the EF-1 alpha promoter in both stem cell lines was considerably higher compared to the CMV promoter. This is mainly due to a considerably lower rate of silencing of the EF-1 alpha promoter in stem cells compared to the CMV promoter as published (Wang, et al. (2008) Stem Cells Dev 17:279–289).

Required Products

Cat. # Product Size Price License Quantity Details
631275 Lenti-X™ Packaging Single Shots (VSV-G) 16 Rxns USD $1071.00

License Statement

ID Number  
63 Use of this product is covered by one or more of the following U.S. Patent Nos. and corresponding patent claims outside the U.S.: 8,562,966, 8,557,231. This product is intended for research purposes only. It may not be used for (i) any human or veterinary use, including without limitation therapeutic and prophylactic use, (ii) any clinical use, including without limitation diagnostic use, (iii) screening of chemical and/or biological compounds for the identification of pharmaceutically active agents (including but not limited to screening of small molecules), target validation, preclinical testing services, or drug development. Any use of this product for any of the above mentioned purposes requires a license from the Massachusetts Institute of Technology.
259 This Product is protected by one or more patents from the family consisting of: JP6454352 and any corresponding patents, divisionals, continuations, patent applications and foreign filings sharing common priority with the same family.

Lenti-X Packaging Single Shots (VSV-G) provide an extremely simple and consistent one-step method for producing high-titer lentivirus. No additional transfection reagent is needed because Lenti-X Packaging Single Shots (VSV-G) consist of pre-aliquoted, lyophilized, single tubes of Xfect Transfection Reagent premixed with an optimized formulation of VSV-G pseudotyped Lenti-X lentiviral packaging plasmids. High-titer virus is produced by simply reconstituting this mixture with your lentiviral vector of choice in sterile water and adding it to 293T cells, e.g., Lenti-X 293T Cells (Cat. # 632180), in a 10 cm dish.

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The Lenti-X Packaging Single Shots (VSV-G) protocol

The Lenti-X Packaging Single Shots (VSV-G) protocol
The Lenti-X Packaging Single Shots (VSV-G) protocol.

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Consistent, high-efficiency transfections lead to high titers

Consistent, high-efficiency transfections lead to high titers

Consistent, high-efficiency transfections lead to high titers. A lentiviral vector containing the ZsGreen1 gene was packaged according to the Lenti-X single shots protocol in four independent experiments. Briefly, 7 µg of pLVX-ZsGreen1 plasmid was added to each of four Lenti-X single shots; the tubes were vortexed for 20 sec and incubated at room temperature for 10 min. Then, the mixture was added to cultured Lenti-X 293T cells that were approximately 80% confluent. 48 hours after transfection, the cells were imaged by fluorescence microscopy (Panel A, top) and light microscopy (Panel A, bottom). After images were taken, the supernatant was harvested and used infect HT1080 cells for titer determination (Panel B, IFU/ml).

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High-titer virus was produced regardless of the lentiviral vector backbone with Lenti-X packaging single shots

High-titer virus was produced regardless of the lentiviral vector backbone with Lenti-X packaging single shots

High-titer virus was produced regardless of the lentiviral vector backbone with Lenti-X packaging single shots. A CMV ZsGreen1 expression cassette was cloned into several lentiviral vector backbones. These vectors were then packaged into lentivirus using the Lenti-X packaging single shots following the provided protocol. Briefly, 7 µg of pLVX-ZsGreen1 plasmid was added to each of four Lenti-X single shots, and the tubes were vortexed for 20 sec and incubated at room temperature for 10 min. Then, the mixture was added to cultured Lenti-X 293T cells that were approximately 80% confluent. After 48 hours, titer was determined using several methods. To determine infectivity, the supernatant was harvested and used to infect HT1080 cells (Flow Cytometry). Harvested viral supernatants were also analyzed by RT-PCR to quantify viral genome copies (qRT-PCR, Lenti-X qRT-PCR Titration Kit), ELISA to measure p24 (p24 ELISA, Lenti-X p24 Rapid Titer Kit), and by a rapid lentiviral detection method (Lenti-X GoStix).

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A comparison of fourth- and third-generation lentiviral packaging systems

A comparison of fourth- and third-generation lentiviral packaging systems

A comparison of fourth- and third-generation lentiviral packaging systems . Our Lenti-X Packaging single shots utilize a packaging system that consists of five separate components (Panel A), mixed in proprietary proportions for optimized packaging activity. The separation of the gag, pol, and env genes effectively reduces the incidence of RCL (Wu et al., 2000). High levels of expression of essential viral components are driven by the Tet-Off and Tat transactivators, which induce a cascade of expression that results in high titers of lentivirus. The pol gene is fused to vpr to ensure transport of the reverse transcriptase/integrase protein into the recombinant lentiviral particle. Not all vector elements are shown. Other 3rd generation lentiviral packaging systems (Panel B) generate lower titers, do not contain separate gag and pol sequences, and do not use a transactivation cascade mechanism.

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631275: Lenti-X Packaging Single Shots (VSV-G)

631275: Lenti-X Packaging Single Shots (VSV-G)
632180 Lenti-X™ 293T Cell Line 1 mL USD $422.00

License Statement

ID Number  
406 This product is the subject of a technology license agreement for internal research use only. Use of this product other than for research use may require additional licenses. Information on license restrictions or for uses other than research may be obtained by contacting licensing@takarabio.com.

The Lenti-X 293T Cell Line is a subclone of the transformed human embryonic kidney cell line, HEK 293, which is highly transfectable and supports high levels of viral protein expression. When transfected with Lenti-X Packaging Single Shots and a lentiviral vector, these cells are capable of producing lentiviral titers as high as >108 ifu/ml, as determined by flow cytometry. The cell line also constitutively expresses the simian virus 40 (SV40) large T antigen.

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632180: Lenti-X 293T Cell Line

632180: Lenti-X 293T Cell Line

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Clontech's lentiviral packaging system (Panel A) and a lentiviral packaging system from a leading competitor (Panel B) were each used to generate viral supernatants from their respective lentiviral system vectors that were engineered to express the ZsGreen1 fluorescent protein

Clontech's lentiviral packaging system (Panel A) and a lentiviral packaging system from a leading competitor (Panel B) were each used to generate viral supernatants from their respective lentiviral system vectors that were engineered to express the ZsGreen1 fluorescent protein
Clontech's lentiviral packaging system (Panel A) and a lentiviral packaging system from a leading competitor (Panel B) were each used to generate viral supernatants from their respective lentiviral system vectors that were engineered to express the ZsGreen1 fluorescent protein. As little as 10 µl of supernatant from Lenti-X transduced the majority of these HeLa cells, whereas 10 µl of supernatant from the other system transduced only a small percentage of the cells. Transduced cells were quantified by flow cytometry.

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Transduction of neural progenitor cells by Lenti-X lentivirus

Transduction of neural progenitor cells by Lenti-X lentivirus

Transduction of neural progenitor cells by Lenti-X lentivirus. Recombinant lentivirus for expressing ZsGreen1 was produced using Lenti-X virus and used to transduce normal human neural progenitor cells. A single transduced cell is shown under phase contrast microscopy (Panel A) and fluorescence microscopy (Panel B).

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High-titer lentivirus production

High-titer lentivirus production

High-titer lentivirus production. Lenti-X 293T cells were transduced with the indicated volumes (µl) of lentiviral packaging supernatant generated with the Lenti-X Expression System and then selected with puromycin for 9 days to allow the formation of the resistant colonies, which were then stained with crystal violet.

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293T cell line for higher titers

293T cell line for higher titers

293T cell line for higher titers. We used our fourth-generation lentiviral packaging system and one of our pLVX-lentiviral vectors to compare the virus production of the Lenti-X 293T Cell Line to that of two other commonly used HEK 293-based cell lines. Lenti-X 293T cells clearly outperformed the other cell lines—producing over 6X more virus than 293FT cells and up to 30X more virus than the parental HEK 293 cell line.

631976 pEF1alpha-IRES-ZsGreen1 Vector 10 ug USD $579.00

pEF1α-IRES-ZsGreen1 is a bicistronic mammalian expression vector that allows the simultaneous, constitutive expression of a protein of interest and the green fluorescent protein ZsGreen1. Constitutive expression of the bicistronic transcript is driven by the human elongation factor 1 alpha (EF1α) promoter.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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631976: pEF1alpha-IRES-ZsGreen1 Vector

631976: pEF1alpha-IRES-ZsGreen1 Vector

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Map schematic of the plasmid choices that are available carrying the EF1-alpha promoter

Map schematic of the plasmid choices that are available carrying the EF1-alpha promoter

Map schematic of the plasmid choices that are available carrying the EF1-alpha promoter. IRES: internal ribosome entry sequence; FP1: fluorescent protein (AcGFP1, DsRed-Monomer, or mCherry); FP2: fluorescent protein (mCherry or ZsGreen1); MCS: multiple cloning site.

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Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter. The mouse embryonic stem cell lines E14 (Panel A) and D3 (Panel B) were transduced by Lenti-X lentivirus, expressing AcGFP1 either under the control of the CMV promoter or the Elongation factor alpha (EF-1 alpha) promoter. The expression level of AcGFP1 in infected cells five days postinfection was monitored by FACS analysis using the FL1 channel. The expression of AcGFP1 driven by the EF-1 alpha promoter in both stem cell lines was considerably higher compared to the CMV promoter. This is mainly due to a considerably lower rate of silencing of the EF-1 alpha promoter in stem cells compared to the CMV promoter as published (Wang, et al. (2008) Stem Cells Dev 17:279–289).

632428 pZsGreen1-DR Vector 20 ug USD $615.00

pZsGreen1-DR is a promoterless vector that encodes a destabilized variant of Zoanthus sp. green fluorescent protein, ZsGreen1. ZsGreen1-DR contains a PEST degradation domain at its C-terminus, so it is rapidly turned over in mammalian cell culture. Because of its short half-life, ZsGreen1-DR is well suited for monitoring the activity of promoters and promoter/enhancer combinations cloned into the multiple cloning site, located upstream of the human codon-optimized ZsGreen1-DR gene. Without the addition of a functional promoter, this vector will not express ZsGreen1-DR.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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Vector map for destabilized fluorescent protein vectors pDsRed-Express-DR, pHcRed1-DR, and pZsGreen1-DR

Vector map for destabilized fluorescent protein vectors pDsRed-Express-DR, pHcRed1-DR, and pZsGreen1-DR
Vector map for destabilized fluorescent protein vectors pDsRed-Express-DR, pHcRed1-DR, and pZsGreen1-DR. These fluorescent protein variants display rapid turnover rates.

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632428: pZsGreen1-DR Vector

632428: pZsGreen1-DR Vector
632473 pZsGreen1-1 Vector 20 ug USD $615.00

pZsGreen1-1 encodes ZsGreen1, a variant of the Zoanthus sp. green fluorescent protein (ZsGreen1) that has been engineered for brighter fluorescence. The ZsGreen1 coding sequence contains a series of silent base pair changes for optimal expression in mammalian cells. pZsGreen1-1 can be used as a reporter to monitor the activity of promoters and promoter/enhancer combinations cloned into the multiple cloning site (MCS) upstream of the promoterless ZsGreen1 gene. Without the addition of a functional promoter, this vector will not express ZsGreen1.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2, and HcRed1

Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2, and HcRed1

Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2,  and HcRed1.

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Vector map for promoterless fluorescent protein vectors AcGFP1-1, DsRed2-1, DsRed-Express-1, HcRed1-1 and ZsGreen1-1

Vector map for promoterless fluorescent protein vectors AcGFP1-1, DsRed2-1, DsRed-Express-1, HcRed1-1 and ZsGreen1-1

Vector map for promoterless fluorescent protein vectors pAcGFP1-1, pDsRed2-1, pDsRed-Express-1, pHcRed1-1 and pZsGreen1-1.

Selection guides Fluorescent protein quick guide
Selection guides Fluorescent protein vector finder

Overview

  • Exceptional brightness: up to 4X brighter than EGFP
  • ZsGreen1 excitation and emission maxima are 493 nm and 505 nm, respectively
  • We offer both monoclonal and polyclonal ZsGreen1 antibodies, which recognize native and denatured forms of ZsGreen1 fluorescent protein, as well as fusion proteins containing ZsGreen1. They do not cross-react with Aequorea sp. GFPs (AcGFP1, EGFP). 
  • Bicistronic (pIRES-) expression vectors are available
  • Lentiviral (pLVX-) and retroviral (pRetroX-) formats are available

More Information

Applications

  • Whole-cell labeling
  • Promoter-reporter studies
  • Transfection controls
References

Matz, M. V. et al. Fluorescent proteins from nonbioluminescent Anthozoa species. Nature Biotechnol. 17, 969–973 (1999).

Additional product information

Please see the product’s Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysisand Kit Components Lists are located under the Documents tab.


Lentiviral-mediated fluorescent protein transduction

Lentiviral fluorescent proteins

We offer a wide array of premade lentivirus particles for whole-cell labeling or for labeling subcellular compartments, such as mitochondria, golgi, membranes, etc.

Whole-cell labeling Subcellular-structure labeling

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Takara Bio USA, Inc. provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, Takara Bio USA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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Takara Bio USA, Inc. provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, Takara Bio USA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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