DsRed-Monomer fluorescent protein

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DsRed-Monomer is soluble when expressed in mammalian cells

DsRed-Monomer is soluble when expressed in mammalian cells. HeLa cells were transfected with pDsRed-Monomer-N1 and fixed in 4% paraformaldehyde 24 hr post-transfection. DsRed-Monomer fluorescent protein displays an even, consistent, and homogeneous distribution.

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AcGFP1 is ideal for multicolor and fluorescence microscopy applications

AcGFP1 is ideal for multicolor and fluorescence microscopy applications. AcGFP1 and DsRed2 protein fusions were transiently transfected and visualized by fluorescence microscopy. Panel A. pAcGFP1-Mito (mitochondria) and pDsRed2-Nuc (nucleus) in HEK 293 cells. Panel B. pAcGFP1-Golgi (Golgi apparatus) and pDsRed2-Nuc (nucleus) in HEK 293 cells.

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Organelles targeted by the subcellular localization vectors

Organelles targeted by the subcellular localization vectors.

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Subcellular localization vectors encode fusions of fluorescent proteins with localization signals or subcellular structural proteins, which target the fluorescent protein to a specific organelle or subcellular structure

Subcellular localization vectors encode fusions of fluorescent proteins with localization signals or subcellular structural proteins, which target the fluorescent protein to a specific organelle or subcellular structure. The vectors are available in a variety of organelle- and cytoskeleton-targeted color variants.

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DsRed-Monomer is a monomeric protein

DsRed-Monomer is a monomeric protein. Panel A. Recombinant DsRed-Express and DsRed-Monomer fluorescent proteins (100 μg) were analyzed by FPLC gel filtration chromatography. Overall absorbance (A₂₈₀) and chromophore excitation (A₅₅₇) of the eluted material were monitored simultaneously. DsRed-Monomer elutes from the column at a retention time (39 min) corresponding to a molecular weight of 28 kDa. The calculated molecular weight of DsRed-Monomer is 26.8 kDa. DsRed-Express is a tetrameric protein that elutes at an earlier retention time (33 min) corresponding to a molecular weight of 89 kDa. Panel B. Pseudonative gel analysis of proteins. The oligomeric structure of proteins is preserved during SDS PAGE analysis if samples are kept at 4°C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins (7.5 μg) were separated by SDS PAGE electrophoresis (12% acrylamide). In both the boiled (denatured) and unboiled (nondenatured) samples, DsRed-Monomer fluorescent protein runs as a uniform band of ~30 kDa due to its monomeric structure. The unboiled (nondenatured) DsRed-Express runs at a much higher molecular weight than its boiled (denatured) counterpart due to its tetrameric structure.

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Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1.

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DsRed-Monomer is soluble when expressed in mammalian cells

DsRed-Monomer is soluble when expressed in mammalian cells. HeLa cells were transfected with pDsRed-Monomer-N1 and fixed in 4% paraformaldehyde 24 hr post-transfection. DsRed-Monomer fluorescent protein displays an even, consistent, and homogeneous distribution.

Back

Organelles targeted by the subcellular localization vectors

Organelles targeted by the subcellular localization vectors.

Back

Subcellular localization vectors encode fusions of fluorescent proteins with localization signals or subcellular structural proteins, which target the fluorescent protein to a specific organelle or subcellular structure

Subcellular localization vectors encode fusions of fluorescent proteins with localization signals or subcellular structural proteins, which target the fluorescent protein to a specific organelle or subcellular structure. The vectors are available in a variety of organelle- and cytoskeleton-targeted color variants.

Back

DsRed-Monomer is a monomeric protein

DsRed-Monomer is a monomeric protein. Panel A. Recombinant DsRed-Express and DsRed-Monomer fluorescent proteins (100 μg) were analyzed by FPLC gel filtration chromatography. Overall absorbance (A₂₈₀) and chromophore excitation (A₅₅₇) of the eluted material were monitored simultaneously. DsRed-Monomer elutes from the column at a retention time (39 min) corresponding to a molecular weight of 28 kDa. The calculated molecular weight of DsRed-Monomer is 26.8 kDa. DsRed-Express is a tetrameric protein that elutes at an earlier retention time (33 min) corresponding to a molecular weight of 89 kDa. Panel B. Pseudonative gel analysis of proteins. The oligomeric structure of proteins is preserved during SDS PAGE analysis if samples are kept at 4°C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins (7.5 μg) were separated by SDS PAGE electrophoresis (12% acrylamide). In both the boiled (denatured) and unboiled (nondenatured) samples, DsRed-Monomer fluorescent protein runs as a uniform band of ~30 kDa due to its monomeric structure. The unboiled (nondenatured) DsRed-Express runs at a much higher molecular weight than its boiled (denatured) counterpart due to its tetrameric structure.

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Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1.

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DsRed-Monomer is soluble when expressed in mammalian cells

DsRed-Monomer is soluble when expressed in mammalian cells. HeLa cells were transfected with pDsRed-Monomer-N1 and fixed in 4% paraformaldehyde 24 hr post-transfection. DsRed-Monomer fluorescent protein displays an even, consistent, and homogeneous distribution.

Back

Organelles targeted by the subcellular localization vectors

Organelles targeted by the subcellular localization vectors.

Back

Subcellular localization vectors encode fusions of fluorescent proteins with localization signals or subcellular structural proteins, which target the fluorescent protein to a specific organelle or subcellular structure

Subcellular localization vectors encode fusions of fluorescent proteins with localization signals or subcellular structural proteins, which target the fluorescent protein to a specific organelle or subcellular structure. The vectors are available in a variety of organelle- and cytoskeleton-targeted color variants.

Back

DsRed-Monomer is a monomeric protein

DsRed-Monomer is a monomeric protein. Panel A. Recombinant DsRed-Express and DsRed-Monomer fluorescent proteins (100 μg) were analyzed by FPLC gel filtration chromatography. Overall absorbance (A₂₈₀) and chromophore excitation (A₅₅₇) of the eluted material were monitored simultaneously. DsRed-Monomer elutes from the column at a retention time (39 min) corresponding to a molecular weight of 28 kDa. The calculated molecular weight of DsRed-Monomer is 26.8 kDa. DsRed-Express is a tetrameric protein that elutes at an earlier retention time (33 min) corresponding to a molecular weight of 89 kDa. Panel B. Pseudonative gel analysis of proteins. The oligomeric structure of proteins is preserved during SDS PAGE analysis if samples are kept at 4°C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins (7.5 μg) were separated by SDS PAGE electrophoresis (12% acrylamide). In both the boiled (denatured) and unboiled (nondenatured) samples, DsRed-Monomer fluorescent protein runs as a uniform band of ~30 kDa due to its monomeric structure. The unboiled (nondenatured) DsRed-Express runs at a much higher molecular weight than its boiled (denatured) counterpart due to its tetrameric structure.

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Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1.

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Retroviral vectors expressing AcGFP1 and DsRed-Monomer fluorescent protein fusions

Retroviral vectors expressing AcGFP1 and DsRed-Monomer fluorescent protein fusions. pRetroQ-DsRed-Monomer-Golgi and pRetroQ-AcGFP1-Tubulin constructs were transfected individually into GP2-293 cells, and the VSV-G pseudotyped virus was harvested 48 hr posttransfection. These viral stocks were then used to coinfect HeLa cells, and expression was visualized 48 hr postinfection by fluorescent microscopy.

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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).

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Retroviral vectors expressing AcGFP1 and DsRed-Monomer fluorescent protein fusions

Retroviral vectors expressing AcGFP1 and DsRed-Monomer fluorescent protein fusions. pRetroQ-DsRed-Monomer-Golgi and pRetroQ-AcGFP1-Tubulin constructs were transfected individually into GP2-293 cells, and the VSV-G pseudotyped virus was harvested 48 hr posttransfection. These viral stocks were then used to coinfect HeLa cells, and expression was visualized 48 hr postinfection by fluorescent microscopy.

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DsRed-Monomer is soluble when expressed in mammalian cells

DsRed-Monomer is soluble when expressed in mammalian cells. HeLa cells were transfected with pDsRed-Monomer-N1 and fixed in 4% paraformaldehyde 24 hr post-transfection. DsRed-Monomer fluorescent protein displays an even, consistent, and homogeneous distribution.

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Cloning of PCR-amplified alpha-actinin directly into four different In-Fusion Ready prelinearized DsRed-Monomer and AcGFP1 vectors

Cloning of PCR-amplified alpha-actinin directly into four different In-Fusion Ready prelinearized DsRed-Monomer and AcGFP1 vectors. The gene for alpha-actinin (1,600 bp) was amplified by PCR and immediately cloned into four different prelinearized DsRed-Monomer N1/C1 and AcGFP1 N1/C1 vectors using the In-Fusion cloning method. All four recombinant vectors were transfected into HeLa cells using a lipid-based transfection agent. 36 hr posttransfection, cells were fixed using 4% paraformaldehyde and visualized using a Zeiss Axioscope fluorescence microscope. Panel A. Alpha-Actinin-AcGFP1-C1. Panel B. Alpha-Actinin-AcGFP-N1. Panel C. Alpha-Actinin-DsRed-Monomer-C1. Panel D. Alpha-Actinin-DsRed-Monomer-N1.

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DsRed-Monomer is soluble when expressed in mammalian cells

DsRed-Monomer is soluble when expressed in mammalian cells. HeLa cells were transfected with pDsRed-Monomer-N1 and fixed in 4% paraformaldehyde 24 hr post-transfection. DsRed-Monomer fluorescent protein displays an even, consistent, and homogeneous distribution.

Back

DsRed-Monomer is a monomeric protein

DsRed-Monomer is a monomeric protein. Panel A. Recombinant DsRed-Express and DsRed-Monomer fluorescent proteins (100 μg) were analyzed by FPLC gel filtration chromatography. Overall absorbance (A₂₈₀) and chromophore excitation (A₅₅₇) of the eluted material were monitored simultaneously. DsRed-Monomer elutes from the column at a retention time (39 min) corresponding to a molecular weight of 28 kDa. The calculated molecular weight of DsRed-Monomer is 26.8 kDa. DsRed-Express is a tetrameric protein that elutes at an earlier retention time (33 min) corresponding to a molecular weight of 89 kDa. Panel B. Pseudonative gel analysis of proteins. The oligomeric structure of proteins is preserved during SDS PAGE analysis if samples are kept at 4°C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins (7.5 μg) were separated by SDS PAGE electrophoresis (12% acrylamide). In both the boiled (denatured) and unboiled (nondenatured) samples, DsRed-Monomer fluorescent protein runs as a uniform band of ~30 kDa due to its monomeric structure. The unboiled (nondenatured) DsRed-Express runs at a much higher molecular weight than its boiled (denatured) counterpart due to its tetrameric structure.

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Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1.

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Cloning of PCR-amplified alpha-actinin directly into four different In-Fusion Ready prelinearized DsRed-Monomer and AcGFP1 vectors

Cloning of PCR-amplified alpha-actinin directly into four different In-Fusion Ready prelinearized DsRed-Monomer and AcGFP1 vectors. The gene for alpha-actinin (1,600 bp) was amplified by PCR and immediately cloned into four different prelinearized DsRed-Monomer N1/C1 and AcGFP1 N1/C1 vectors using the In-Fusion cloning method. All four recombinant vectors were transfected into HeLa cells using a lipid-based transfection agent. 36 hr posttransfection, cells were fixed using 4% paraformaldehyde and visualized using a Zeiss Axioscope fluorescence microscope. Panel A. Alpha-Actinin-AcGFP1-C1. Panel B. Alpha-Actinin-AcGFP-N1. Panel C. Alpha-Actinin-DsRed-Monomer-C1. Panel D. Alpha-Actinin-DsRed-Monomer-N1.

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DsRed-Monomer is soluble when expressed in mammalian cells

DsRed-Monomer is soluble when expressed in mammalian cells. HeLa cells were transfected with pDsRed-Monomer-N1 and fixed in 4% paraformaldehyde 24 hr post-transfection. DsRed-Monomer fluorescent protein displays an even, consistent, and homogeneous distribution.

Back

DsRed-Monomer is a monomeric protein

DsRed-Monomer is a monomeric protein. Panel A. Recombinant DsRed-Express and DsRed-Monomer fluorescent proteins (100 μg) were analyzed by FPLC gel filtration chromatography. Overall absorbance (A₂₈₀) and chromophore excitation (A₅₅₇) of the eluted material were monitored simultaneously. DsRed-Monomer elutes from the column at a retention time (39 min) corresponding to a molecular weight of 28 kDa. The calculated molecular weight of DsRed-Monomer is 26.8 kDa. DsRed-Express is a tetrameric protein that elutes at an earlier retention time (33 min) corresponding to a molecular weight of 89 kDa. Panel B. Pseudonative gel analysis of proteins. The oligomeric structure of proteins is preserved during SDS PAGE analysis if samples are kept at 4°C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins (7.5 μg) were separated by SDS PAGE electrophoresis (12% acrylamide). In both the boiled (denatured) and unboiled (nondenatured) samples, DsRed-Monomer fluorescent protein runs as a uniform band of ~30 kDa due to its monomeric structure. The unboiled (nondenatured) DsRed-Express runs at a much higher molecular weight than its boiled (denatured) counterpart due to its tetrameric structure.

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Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1.

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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.

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Hygromycin DsRed-Monomer and AcGFP1 expression vectors

Hygromycin DsRed-Monomer and AcGFP1 expression vectors. Vector maps of our DsRed-Monomer and pAcGFP1 Hyg-N1 (Panel A) and Hyg-C1 (Panel B) mammalian expression vectors for fusion applications. Each vector contains a hygromycin selection cassette. These vectors can be used together with our standard N1/C1 vectors containing the neomycin selection cassette to establish double stable cell lines expressing two different Living Colors proteins.

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DsRed-Monomer is soluble when expressed in mammalian cells

DsRed-Monomer is soluble when expressed in mammalian cells. HeLa cells were transfected with pDsRed-Monomer-N1 and fixed in 4% paraformaldehyde 24 hr post-transfection. DsRed-Monomer fluorescent protein displays an even, consistent, and homogeneous distribution.

Back

DsRed-Monomer is a monomeric protein

DsRed-Monomer is a monomeric protein. Panel A. Recombinant DsRed-Express and DsRed-Monomer fluorescent proteins (100 μg) were analyzed by FPLC gel filtration chromatography. Overall absorbance (A₂₈₀) and chromophore excitation (A₅₅₇) of the eluted material were monitored simultaneously. DsRed-Monomer elutes from the column at a retention time (39 min) corresponding to a molecular weight of 28 kDa. The calculated molecular weight of DsRed-Monomer is 26.8 kDa. DsRed-Express is a tetrameric protein that elutes at an earlier retention time (33 min) corresponding to a molecular weight of 89 kDa. Panel B. Pseudonative gel analysis of proteins. The oligomeric structure of proteins is preserved during SDS PAGE analysis if samples are kept at 4°C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins (7.5 μg) were separated by SDS PAGE electrophoresis (12% acrylamide). In both the boiled (denatured) and unboiled (nondenatured) samples, DsRed-Monomer fluorescent protein runs as a uniform band of ~30 kDa due to its monomeric structure. The unboiled (nondenatured) DsRed-Express runs at a much higher molecular weight than its boiled (denatured) counterpart due to its tetrameric structure.

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Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1.

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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.

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Hygromycin DsRed-Monomer and AcGFP1 expression vectors

Hygromycin DsRed-Monomer and AcGFP1 expression vectors. Vector maps of our DsRed-Monomer and pAcGFP1 Hyg-N1 (Panel A) and Hyg-C1 (Panel B) mammalian expression vectors for fusion applications. Each vector contains a hygromycin selection cassette. These vectors can be used together with our standard N1/C1 vectors containing the neomycin selection cassette to establish double stable cell lines expressing two different Living Colors proteins.

Back

DsRed-Monomer is soluble when expressed in mammalian cells

DsRed-Monomer is soluble when expressed in mammalian cells. HeLa cells were transfected with pDsRed-Monomer-N1 and fixed in 4% paraformaldehyde 24 hr post-transfection. DsRed-Monomer fluorescent protein displays an even, consistent, and homogeneous distribution.

Back

DsRed-Monomer is a monomeric protein

DsRed-Monomer is a monomeric protein. Panel A. Recombinant DsRed-Express and DsRed-Monomer fluorescent proteins (100 μg) were analyzed by FPLC gel filtration chromatography. Overall absorbance (A₂₈₀) and chromophore excitation (A₅₅₇) of the eluted material were monitored simultaneously. DsRed-Monomer elutes from the column at a retention time (39 min) corresponding to a molecular weight of 28 kDa. The calculated molecular weight of DsRed-Monomer is 26.8 kDa. DsRed-Express is a tetrameric protein that elutes at an earlier retention time (33 min) corresponding to a molecular weight of 89 kDa. Panel B. Pseudonative gel analysis of proteins. The oligomeric structure of proteins is preserved during SDS PAGE analysis if samples are kept at 4°C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins (7.5 μg) were separated by SDS PAGE electrophoresis (12% acrylamide). In both the boiled (denatured) and unboiled (nondenatured) samples, DsRed-Monomer fluorescent protein runs as a uniform band of ~30 kDa due to its monomeric structure. The unboiled (nondenatured) DsRed-Express runs at a much higher molecular weight than its boiled (denatured) counterpart due to its tetrameric structure.

Back

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1.

Back

AcGFP1 is ideal for multicolor and fluorescence microscopy applications

AcGFP1 is ideal for multicolor and fluorescence microscopy applications. AcGFP1 and DsRed2 protein fusions were transiently transfected and visualized by fluorescence microscopy. Panel A. pAcGFP1-Mito (mitochondria) and pDsRed2-Nuc (nucleus) in HEK 293 cells. Panel B. pAcGFP1-Golgi (Golgi apparatus) and pDsRed2-Nuc (nucleus) in HEK 293 cells.

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Organelles targeted by the subcellular localization vectors

Organelles targeted by the subcellular localization vectors.

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Subcellular localization vectors encode fusions of fluorescent proteins with localization signals or subcellular structural proteins, which target the fluorescent protein to a specific organelle or subcellular structure

Subcellular localization vectors encode fusions of fluorescent proteins with localization signals or subcellular structural proteins, which target the fluorescent protein to a specific organelle or subcellular structure. The vectors are available in a variety of organelle- and cytoskeleton-targeted color variants.

Back

DsRed-Monomer is soluble when expressed in mammalian cells

DsRed-Monomer is soluble when expressed in mammalian cells. HeLa cells were transfected with pDsRed-Monomer-N1 and fixed in 4% paraformaldehyde 24 hr post-transfection. DsRed-Monomer fluorescent protein displays an even, consistent, and homogeneous distribution.

Back

DsRed-Monomer is a monomeric protein

DsRed-Monomer is a monomeric protein. Panel A. Recombinant DsRed-Express and DsRed-Monomer fluorescent proteins (100 μg) were analyzed by FPLC gel filtration chromatography. Overall absorbance (A₂₈₀) and chromophore excitation (A₅₅₇) of the eluted material were monitored simultaneously. DsRed-Monomer elutes from the column at a retention time (39 min) corresponding to a molecular weight of 28 kDa. The calculated molecular weight of DsRed-Monomer is 26.8 kDa. DsRed-Express is a tetrameric protein that elutes at an earlier retention time (33 min) corresponding to a molecular weight of 89 kDa. Panel B. Pseudonative gel analysis of proteins. The oligomeric structure of proteins is preserved during SDS PAGE analysis if samples are kept at 4°C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins (7.5 μg) were separated by SDS PAGE electrophoresis (12% acrylamide). In both the boiled (denatured) and unboiled (nondenatured) samples, DsRed-Monomer fluorescent protein runs as a uniform band of ~30 kDa due to its monomeric structure. The unboiled (nondenatured) DsRed-Express runs at a much higher molecular weight than its boiled (denatured) counterpart due to its tetrameric structure.

Back

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1.

Back

Organelles targeted by the subcellular localization vectors

Organelles targeted by the subcellular localization vectors.

Back

Subcellular localization vectors encode fusions of fluorescent proteins with localization signals or subcellular structural proteins, which target the fluorescent protein to a specific organelle or subcellular structure

Subcellular localization vectors encode fusions of fluorescent proteins with localization signals or subcellular structural proteins, which target the fluorescent protein to a specific organelle or subcellular structure. The vectors are available in a variety of organelle- and cytoskeleton-targeted color variants.

Back

DsRed-Monomer is soluble when expressed in mammalian cells

DsRed-Monomer is soluble when expressed in mammalian cells. HeLa cells were transfected with pDsRed-Monomer-N1 and fixed in 4% paraformaldehyde 24 hr post-transfection. DsRed-Monomer fluorescent protein displays an even, consistent, and homogeneous distribution.

Back

DsRed-Monomer is a monomeric protein

DsRed-Monomer is a monomeric protein. Panel A. Recombinant DsRed-Express and DsRed-Monomer fluorescent proteins (100 μg) were analyzed by FPLC gel filtration chromatography. Overall absorbance (A₂₈₀) and chromophore excitation (A₅₅₇) of the eluted material were monitored simultaneously. DsRed-Monomer elutes from the column at a retention time (39 min) corresponding to a molecular weight of 28 kDa. The calculated molecular weight of DsRed-Monomer is 26.8 kDa. DsRed-Express is a tetrameric protein that elutes at an earlier retention time (33 min) corresponding to a molecular weight of 89 kDa. Panel B. Pseudonative gel analysis of proteins. The oligomeric structure of proteins is preserved during SDS PAGE analysis if samples are kept at 4°C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins (7.5 μg) were separated by SDS PAGE electrophoresis (12% acrylamide). In both the boiled (denatured) and unboiled (nondenatured) samples, DsRed-Monomer fluorescent protein runs as a uniform band of ~30 kDa due to its monomeric structure. The unboiled (nondenatured) DsRed-Express runs at a much higher molecular weight than its boiled (denatured) counterpart due to its tetrameric structure.

Back

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1.

Back

Organelles targeted by the subcellular localization vectors

Organelles targeted by the subcellular localization vectors.

Back

Subcellular localization vectors encode fusions of fluorescent proteins with localization signals or subcellular structural proteins, which target the fluorescent protein to a specific organelle or subcellular structure

Subcellular localization vectors encode fusions of fluorescent proteins with localization signals or subcellular structural proteins, which target the fluorescent protein to a specific organelle or subcellular structure. The vectors are available in a variety of organelle- and cytoskeleton-targeted color variants.

Back

DsRed-Monomer is soluble when expressed in mammalian cells

DsRed-Monomer is soluble when expressed in mammalian cells. HeLa cells were transfected with pDsRed-Monomer-N1 and fixed in 4% paraformaldehyde 24 hr post-transfection. DsRed-Monomer fluorescent protein displays an even, consistent, and homogeneous distribution.

Back

DsRed-Monomer is a monomeric protein

DsRed-Monomer is a monomeric protein. Panel A. Recombinant DsRed-Express and DsRed-Monomer fluorescent proteins (100 μg) were analyzed by FPLC gel filtration chromatography. Overall absorbance (A₂₈₀) and chromophore excitation (A₅₅₇) of the eluted material were monitored simultaneously. DsRed-Monomer elutes from the column at a retention time (39 min) corresponding to a molecular weight of 28 kDa. The calculated molecular weight of DsRed-Monomer is 26.8 kDa. DsRed-Express is a tetrameric protein that elutes at an earlier retention time (33 min) corresponding to a molecular weight of 89 kDa. Panel B. Pseudonative gel analysis of proteins. The oligomeric structure of proteins is preserved during SDS PAGE analysis if samples are kept at 4°C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins (7.5 μg) were separated by SDS PAGE electrophoresis (12% acrylamide). In both the boiled (denatured) and unboiled (nondenatured) samples, DsRed-Monomer fluorescent protein runs as a uniform band of ~30 kDa due to its monomeric structure. The unboiled (nondenatured) DsRed-Express runs at a much higher molecular weight than its boiled (denatured) counterpart due to its tetrameric structure.

Back

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1.

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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.

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DsRed-Monomer is soluble when expressed in mammalian cells

DsRed-Monomer is soluble when expressed in mammalian cells. HeLa cells were transfected with pDsRed-Monomer-N1 and fixed in 4% paraformaldehyde 24 hr post-transfection. DsRed-Monomer fluorescent protein displays an even, consistent, and homogeneous distribution.

Back

DsRed-Monomer is a monomeric protein

DsRed-Monomer is a monomeric protein. Panel A. Recombinant DsRed-Express and DsRed-Monomer fluorescent proteins (100 μg) were analyzed by FPLC gel filtration chromatography. Overall absorbance (A₂₈₀) and chromophore excitation (A₅₅₇) of the eluted material were monitored simultaneously. DsRed-Monomer elutes from the column at a retention time (39 min) corresponding to a molecular weight of 28 kDa. The calculated molecular weight of DsRed-Monomer is 26.8 kDa. DsRed-Express is a tetrameric protein that elutes at an earlier retention time (33 min) corresponding to a molecular weight of 89 kDa. Panel B. Pseudonative gel analysis of proteins. The oligomeric structure of proteins is preserved during SDS PAGE analysis if samples are kept at 4°C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins (7.5 μg) were separated by SDS PAGE electrophoresis (12% acrylamide). In both the boiled (denatured) and unboiled (nondenatured) samples, DsRed-Monomer fluorescent protein runs as a uniform band of ~30 kDa due to its monomeric structure. The unboiled (nondenatured) DsRed-Express runs at a much higher molecular weight than its boiled (denatured) counterpart due to its tetrameric structure.

Back

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.

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Hygromycin DsRed-Monomer and AcGFP1 expression vectors

Hygromycin DsRed-Monomer and AcGFP1 expression vectors. Vector maps of our DsRed-Monomer and pAcGFP1 Hyg-N1 (Panel A) and Hyg-C1 (Panel B) mammalian expression vectors for fusion applications. Each vector contains a hygromycin selection cassette. These vectors can be used together with our standard N1/C1 vectors containing the neomycin selection cassette to establish double stable cell lines expressing two different Living Colors proteins.

Back

DsRed-Monomer is soluble when expressed in mammalian cells

DsRed-Monomer is soluble when expressed in mammalian cells. HeLa cells were transfected with pDsRed-Monomer-N1 and fixed in 4% paraformaldehyde 24 hr post-transfection. DsRed-Monomer fluorescent protein displays an even, consistent, and homogeneous distribution.

Back

DsRed-Monomer is a monomeric protein

DsRed-Monomer is a monomeric protein. Panel A. Recombinant DsRed-Express and DsRed-Monomer fluorescent proteins (100 μg) were analyzed by FPLC gel filtration chromatography. Overall absorbance (A₂₈₀) and chromophore excitation (A₅₅₇) of the eluted material were monitored simultaneously. DsRed-Monomer elutes from the column at a retention time (39 min) corresponding to a molecular weight of 28 kDa. The calculated molecular weight of DsRed-Monomer is 26.8 kDa. DsRed-Express is a tetrameric protein that elutes at an earlier retention time (33 min) corresponding to a molecular weight of 89 kDa. Panel B. Pseudonative gel analysis of proteins. The oligomeric structure of proteins is preserved during SDS PAGE analysis if samples are kept at 4°C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins (7.5 μg) were separated by SDS PAGE electrophoresis (12% acrylamide). In both the boiled (denatured) and unboiled (nondenatured) samples, DsRed-Monomer fluorescent protein runs as a uniform band of ~30 kDa due to its monomeric structure. The unboiled (nondenatured) DsRed-Express runs at a much higher molecular weight than its boiled (denatured) counterpart due to its tetrameric structure.

Back

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1.

Back

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.

Back

Hygromycin DsRed-Monomer and AcGFP1 expression vectors

Hygromycin DsRed-Monomer and AcGFP1 expression vectors. Vector maps of our DsRed-Monomer and pAcGFP1 Hyg-N1 (Panel A) and Hyg-C1 (Panel B) mammalian expression vectors for fusion applications. Each vector contains a hygromycin selection cassette. These vectors can be used together with our standard N1/C1 vectors containing the neomycin selection cassette to establish double stable cell lines expressing two different Living Colors proteins.

Back

DsRed-Monomer is soluble when expressed in mammalian cells

DsRed-Monomer is soluble when expressed in mammalian cells. HeLa cells were transfected with pDsRed-Monomer-N1 and fixed in 4% paraformaldehyde 24 hr post-transfection. DsRed-Monomer fluorescent protein displays an even, consistent, and homogeneous distribution.

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DsRed-Monomer is a monomeric protein

DsRed-Monomer is a monomeric protein. Panel A. Recombinant DsRed-Express and DsRed-Monomer fluorescent proteins (100 μg) were analyzed by FPLC gel filtration chromatography. Overall absorbance (A₂₈₀) and chromophore excitation (A₅₅₇) of the eluted material were monitored simultaneously. DsRed-Monomer elutes from the column at a retention time (39 min) corresponding to a molecular weight of 28 kDa. The calculated molecular weight of DsRed-Monomer is 26.8 kDa. DsRed-Express is a tetrameric protein that elutes at an earlier retention time (33 min) corresponding to a molecular weight of 89 kDa. Panel B. Pseudonative gel analysis of proteins. The oligomeric structure of proteins is preserved during SDS PAGE analysis if samples are kept at 4°C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins (7.5 μg) were separated by SDS PAGE electrophoresis (12% acrylamide). In both the boiled (denatured) and unboiled (nondenatured) samples, DsRed-Monomer fluorescent protein runs as a uniform band of ~30 kDa due to its monomeric structure. The unboiled (nondenatured) DsRed-Express runs at a much higher molecular weight than its boiled (denatured) counterpart due to its tetrameric structure.

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Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1.

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Lentiviral vector delivery of fluorescently labeled protein expression to human neural progenitor cells

Lentiviral vector delivery of fluorescently labeled protein expression to human neural progenitor cells. Neurospheres were allowed to differentiate on laminin-coated plates and then coinfected with equivalent amounts of Lenti-X viruses produced from a pLVX-AcGFP1-Actin vector and a pLVX-DsRed-Monomer-Nuc vector. AcGFP1-labeled actin allows clear visualization of the cytoskeleton (green), while DsRed-Monomer-labeled nuclei (red) are also visible.

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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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Lentiviral vector delivery of fluorescently labeled protein expression to human neural progenitor cells

Lentiviral vector delivery of fluorescently labeled protein expression to human neural progenitor cells. Neurospheres were allowed to differentiate on laminin-coated plates and then coinfected with equivalent amounts of Lenti-X viruses produced from a pLVX-AcGFP1-Actin vector and a pLVX-DsRed-Monomer-Nuc vector. AcGFP1-labeled actin allows clear visualization of the cytoskeleton (green), while DsRed-Monomer-labeled nuclei (red) are also visible.

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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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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).

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631989: pLVX-EF1alpha-DsRed-Monomer-C1 Vector

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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. 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).

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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. 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. 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).

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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. 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).