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Home › Products › Protein research › Expression vectors & systems › Baculovirus expression system › Baculovirus titration kits

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Baculovirus titration kits

Boxes, bottles, and tubes of the BacPAK Baculovirus Rapid Titer Kit

Accurate baculovirus titering is a vital step in determining the multiplicity of infection (MOI), a critical parameter for achieving optimal and consistent protein yield with baculovirus gene expression. However, traditional methods, such as plaque assays and end-point dilution assays, are laborious and take several days.

Our four BacPAK titration solutions are simple, accurate, and less time-consuming than traditional methods. They detect expression of either the baculoviral gp64 protein or p35 protein for quantification. The gp64 protein is expressed during the later stages of baculoviral infection and is only present in viruses capable of infection, while the p35 protein is expressed during the early stages of infection and can be correlated to functional titer.

Trying to decide which baculovirus titration product is right for your workflow? Check out the comparison chart below.

Cat. # Product Size Price License Quantity Details
631479 BacPAK™ Baculovirus GoStix™ Plus 20 Tests USD $647.00

BacPAK Baculovirus GoStix Plus provide a rapid, simple, and effective method for instantly quantifying baculovirus in cell culture supernatants. The test involves applying only 20 µl of precious viral supernatant to a GoStix cassette and waiting 10 minutes for the appearance of test and control bands that indicate the presence of baculoviral gp64. The results on the cassette can then be analyzed using our free smartphone app, which quantifies titer by comparing the intensities of the test and control bands.

The quantitative output from the GoStix App (referred to as a GoStix Value, or GV) can be used to compare virus amounts between different preparations in a manner similar to, but far easier and quicker than, an ELISA. It also allows for the calculation of actual PFU/ml using a reference virus.

Cat # 631479 contains BacPAK Baculovirus GoStix Plus cassettes, Chase Buffer, and gp64 Control.

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.

Documents Components Image Data

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Comparison of baculovirus titration methods

Comparison of baculovirus titration methods

Comparison of baculovirus titration methods. A strong correlation was observed between the physical titration methods (GoStix and qPCR) and the functional methods that require infection (ELISA and plaque staining). The time required to obtain results for each method is summarized in the table.

ZsGreen1-expressing baculovirus was generated using the BacPAK Baculovirus Expression System (Cat. # 631402), following the manufacturer’s protocol. Baculovirus supernatant was harvested from several plaques and amplified in fresh Sf21 cells (P1) for four days.

The baculovirus supernatant from a single clone was analyzed using two functional titration methods, the BacPAK Baculovirus Rapid Titer Kit (Cat. # 631406) and the BacPAK p35 ELISA Kit (Single Wash) (Cat. # 631477), as well as two physical titration methods, the BacPAK qPCR Titration Kit (Cat. # 631414) and the BacPAK Baculovirus GoStix Plus (Cat. # 631479).

Infectious units (IFU) and GoStix results from Clone 11 (data not shown) were used as a reference with our online calculator to convert the units from the different methods into IFU for Clone 4 (shown here). The formula used for these conversions is available in the user manual.

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Timelines associated with most commonly used methods of baculoviral vector titration

Timelines associated with most commonly used methods of baculoviral vector titration

Timelines associated with most commonly used methods of baculoviral vector titration. BacPAK GoStix Plus and ELISA methods are significantly faster than traditional methods and do not require the special equipment of qPCR- and FACS-based methods. FACS = fluorescence-activated cell sorting.

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BacPak GoStix and ELISA baculovirus titration methods can be used at various stages in your baculovirus workflow

BacPak GoStix and ELISA baculovirus titration methods can be used at various stages in your baculovirus workflow

BacPAK GoStix Plus and ELISA baculovirus titration methods can be used at various stages in your baculovirus workflow. BacPAK Baculovirus GoStix Plus (A) can be used for quick titer checks in stages such as clone screening or starting from a stored viral stock for downstream production. BacPAK p35 ELISA Kit (Single Wash) (B) can be used to determine functional titers in steps such as baculovirus titer normalization and expression optimization. You can also use these two products in other stages of your workflow.

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631479: BacPAK Baculovirus GoStix Plus

631479: BacPAK Baculovirus GoStix Plus
631477 BacPAK™ p35 ELISA Kit (Single Wash) 96 Rxns USD $696.00

BacPAK p35 ELISA Kit (Single Wash) enables you to determine functional baculovirus titers based on the expression of baculoviral p35. The ELISA protocol has been developed to reduce the number of steps, which minimizes hands-on time. The entire protocol can be completed in 20 hours, including an overnight infection step. However, only 2 hrs of hands-on time is required. The resulting p35 titer value can easily be converted to IFU/ml with our Baculovirus p35-to-IFU calculator.

Cat. # 631477 contains an Anti-p35 Coated Plate of 96 wells (available as 12 x 8-well strips), p35 Control, Lysis Buffer, p35 Buffer, Anti-p35 (HRP Conjugate), Washing Buffer (20X), TMB Substrate, and Stop Solution.

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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631477: BacPAK p35 ELISA Kit (Single Wash)

631477: BacPAK p35 ELISA Kit (Single Wash)

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Comparison of baculovirus titration methods

Comparison of baculovirus titration methods

Comparison of baculovirus titration methods. A strong correlation was observed between the physical titration methods (GoStix and qPCR) and the functional methods that require infection (ELISA and plaque staining). The time required to obtain results for each method is summarized in the table.

ZsGreen1-expressing baculovirus was generated using the BacPAK Baculovirus Expression System (Cat. # 631402), following the manufacturer’s protocol. Baculovirus supernatant was harvested from several plaques and amplified in fresh Sf21 cells (P1) for four days.

The baculovirus supernatant from a single clone was analyzed using two functional titration methods, the BacPAK Baculovirus Rapid Titer Kit (Cat. # 631406) and the BacPAK p35 ELISA Kit (Single Wash) (Cat. # 631477), as well as two physical titration methods, the BacPAK qPCR Titration Kit (Cat. # 631414) and the BacPAK Baculovirus GoStix Plus (Cat. # 631479).

Infectious units (IFU) and GoStix results from Clone 11 (data not shown) were used as a reference with our online calculator to convert the units from the different methods into IFU for Clone 4 (shown here). The formula used for these conversions is available in the user manual.

Back

Timelines associated with most commonly used methods of baculoviral vector titration

Timelines associated with most commonly used methods of baculoviral vector titration

Timelines associated with most commonly used methods of baculoviral vector titration. BacPAK GoStix Plus and ELISA methods are significantly faster than traditional methods and do not require the special equipment of qPCR- and FACS-based methods. FACS = fluorescence-activated cell sorting.

Back

BacPak GoStix and ELISA baculovirus titration methods can be used at various stages in your baculovirus workflow

BacPak GoStix and ELISA baculovirus titration methods can be used at various stages in your baculovirus workflow

BacPAK GoStix Plus and ELISA baculovirus titration methods can be used at various stages in your baculovirus workflow. BacPAK Baculovirus GoStix Plus (A) can be used for quick titer checks in stages such as clone screening or starting from a stored viral stock for downstream production. BacPAK p35 ELISA Kit (Single Wash) (B) can be used to determine functional titers in steps such as baculovirus titer normalization and expression optimization. You can also use these two products in other stages of your workflow.

631406 BacPAK™ Baculovirus Rapid Titer Kit 5 Assays USD $574.00

License Statement

ID Number  
330 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 BacPAK Baculovirus Rapid Titer Kit enables you to determine infectious baculovirus titers based on the expression of baculoviral gp64. The immunological staining protocol can be completed in 2 days.

Cat. # 631406 includes Goat Anti-mouse Antibody/HRP Conjugate, Normal Goat Serum, Mouse gp64 Antibody, Blue Peroxidase Substrate, Methyl Cellulose Overlay, and Resealable Plastic Bags.

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.

Documents Components You May Also Like Image Data

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Identification of infection foci in a lawn of Sf21 cells, using the BacPAK Baculovirus Rapid Titer Kit

Identification of infection foci in a lawn of Sf21 cells, using the BacPAK Baculovirus Rapid Titer Kit
Identification of infection foci in a lawn of Sf21 cells, using the BacPAK Baculovirus Rapid Titer Kit. Cells were immunostained with the gp64 primary antibody and an HRP-conjugated secondary antibody, and visualized by light microscopy. Panel A. Former protocol, with formyl-buffered acetone fixation. Panel B. Improved protocol, with 4% paraformaldehyde fixation.

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The In-Fusion Ready BacPAK Vector Set and Baculovirus Expression System

The In-Fusion Ready BacPAK Vector Set and Baculovirus Expression System
The In-Fusion Ready BacPAK Vector Set and Baculovirus Expression System. A PCR fragment containing your gene of interest is simultaneously and directly cloned into the In-Fusion Ready BacPAK Vector pair to generate N- and C-terminal-tagged constructs. Verified clones are then cotransfected with BacPAK6 viral DNA into insect cells to initiate recombination via the AcMNPV sequences, and replication of recombinant virus. Following virus amplification, infection of insect cell cultures produces recombinant protein, which is then purified by TALON technology.

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Insect cells infected with a recombinant baculovirus express the Aequorea coerulescens green fluorescent protein (AcGFP1)

Insect cells infected with a recombinant baculovirus express the Aequorea coerulescens green fluorescent protein (AcGFP1)
Insect cells infected with a recombinant baculovirus express the Aequorea coerulescens green fluorescent protein (AcGFP1). An In-Fusion Ready BacPAK vector was used to generate a recombinant baculovirus harboring an N-terminal tagged expression construct for the Living Colors fluorescent protein, AcGFP1. Sf9 cells infected with the virus expressed high levels of AcGFP1 and became highly fluorescent. Analysis by flow cytometry revealed that the mean fluorescence intensity of the infected cells was approximately 440-fold greater than that of the uninfected control.

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Insect cells express N- and C-tagged Aequorea coerulescens green fluorescent protein (AcGFP1)

Insect cells express N- and C-tagged Aequorea coerulescens green fluorescent protein (AcGFP1)
Insect cells express N- and C-tagged Aequorea coerulescens green fluorescent protein (AcGFP1). In-Fusion Ready BacPAK vectors were used to generate recombinant baculoviruses harboring N- or C-terminal-tagged AcGFP1 expression constructs. Insect cells (Sf9) infected with either virus express AcGFP1 and thus appear green when viewed by fluorescence microscopy. Panel A. BacPAK-Nterm-6xHN-AcGFP1. Panel B. BacPAK-Cterm- 6xHN-AcGFP1

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Flow chart of the two-day BacPAK Baculovirus Rapid Titer Kit procedure

Flow chart of the two-day BacPAK Baculovirus Rapid Titer Kit procedure
Flow chart of the two-day BacPAK Baculovirus Rapid Titer Kit procedure.

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631406: BacPAK Baculovirus Rapid Titer Kit

631406: BacPAK Baculovirus Rapid Titer Kit
631414 BacPAK™ qPCR Titration Kit 200 Rxns USD $602.00

The BacPAK qPCR Titration Kit rapidly quantifies the virion content of baculoviral supernatants. The copy numbers of viral DNA genomes contained in virus samples are determined by comparing the qPCR threshold cycles (Ct) derived from serial dilutions of purified viral DNA to the Ct values derived from dilutions of a calibrated control template.

Cat # 631414 includes a viral nucleic acid purification kit, calibrated BacPAK DNA Control Template, primers, and all the reagents required to conduct 200 qPCR reactions using either the BacPAK DNA Control Template or purified viral DNA samples.

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.

Documents Components Image Data

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The BacPAK qPCR Titration kit offers a wide dynamic range

The BacPAK qPCR Titration kit offers a wide dynamic range
The BacPAK qPCR Titration kit offers a wide dynamic range. The BacPAK DNA Control Template was serially diluted from 108–103 copies per sample and analyzed with the BacPAK qPCR Titration Kit. The amplification plots (Panel A) show a dynamic range of at least six orders of magnitude with no NTC (No-Template Control) background. The standard curve (Panel B) obtained by plotting the Ct values (determined from the amplification plots in Panel A) against the log of the DNA copy number in each sample, demonstrates a strong linear correlation between the Ct and the DNA copy number (log scale), with R2 = 1.000 and a PCR efficiency of 100.0.07.

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The In-Fusion Ready BacPAK Vector Set and Baculovirus Expression System

The In-Fusion Ready BacPAK Vector Set and Baculovirus Expression System
The In-Fusion Ready BacPAK Vector Set and Baculovirus Expression System. A PCR fragment containing your gene of interest is simultaneously and directly cloned into the In-Fusion Ready BacPAK Vector pair to generate N- and C-terminal-tagged constructs. Verified clones are then cotransfected with BacPAK6 viral DNA into insect cells to initiate recombination via the AcMNPV sequences, and replication of recombinant virus. Following virus amplification, infection of insect cell cultures produces recombinant protein, which is then purified by TALON technology.

Back

Insect cells infected with a recombinant baculovirus express the Aequorea coerulescens green fluorescent protein (AcGFP1)

Insect cells infected with a recombinant baculovirus express the Aequorea coerulescens green fluorescent protein (AcGFP1)
Insect cells infected with a recombinant baculovirus express the Aequorea coerulescens green fluorescent protein (AcGFP1). An In-Fusion Ready BacPAK vector was used to generate a recombinant baculovirus harboring an N-terminal tagged expression construct for the Living Colors fluorescent protein, AcGFP1. Sf9 cells infected with the virus expressed high levels of AcGFP1 and became highly fluorescent. Analysis by flow cytometry revealed that the mean fluorescence intensity of the infected cells was approximately 440-fold greater than that of the uninfected control.

Back

Insect cells express N- and C-tagged Aequorea coerulescens green fluorescent protein (AcGFP1)

Insect cells express N- and C-tagged Aequorea coerulescens green fluorescent protein (AcGFP1)
Insect cells express N- and C-tagged Aequorea coerulescens green fluorescent protein (AcGFP1). In-Fusion Ready BacPAK vectors were used to generate recombinant baculoviruses harboring N- or C-terminal-tagged AcGFP1 expression constructs. Insect cells (Sf9) infected with either virus express AcGFP1 and thus appear green when viewed by fluorescence microscopy. Panel A. BacPAK-Nterm-6xHN-AcGFP1. Panel B. BacPAK-Cterm- 6xHN-AcGFP1

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The BacPAK qPCR Titration Protocol

The BacPAK qPCR Titration Protocol
The BacPAK qPCR Titration Protocol.

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631414: BacPAK qPCR Titration Kit

631414: BacPAK qPCR Titration Kit

p35 IFU calculator tool GoStix Plus-to-IFU calculator
p35 IFU calculator tool Baculovirus p35-to-IFU calculator
p35 IFU calculator tool BacPAK special offer

Overview

Our BacPAK titration solutions allow you to:

  • Shorten baculovirus expression experiments by up to six days
  • Eliminate troublesome plaque assays and end-point dilution assays
  • Use with any AcMNPV-based baculovirus expression systems
  • Harvest, titer, and infect in a single day
BacPAK product comparison chart
BacPAK Baculovirus GoStix Plus BacPAK p35 ELISA Kit (Single Wash) BacPAK Baculovirus Rapid Titer Kit BacPAK qPCR Titration Kit
Method Lateral flow assay ELISA Immunological staining qPCR
Assay time 10 min 20 hr 2 days 4 hr
Titers are based on gp64 expression p35 expression gp64 expression Viral genome content
Sensitivity Low Medium Medium High
Dynamic range 1–30 ng gp64 1 x 105–1 x 107 PFU/ml 1 x 105–1 x 107 PFU/ml 1 x 104–1 x 108 PFU/ml
Advantages/disadvantages Fastest and easiest method; provides accurate physical titer quantification but is low throughput. Yields functional titer and can perform up to 96 samples at one time. Requires an overnight incubation, but little actual hands-on time. Yields physical titer; can perform 96 samples at one time. Potentially more sensitive than other methods, but does not give a readout of functional titer
When to choose this product You want to quickly determine if your baculovirus is ready to harvest and/or you are willing to process samples individually. You want to determine functional titer and want to process multiple samples at once. You want to base your titers on gp64 measurement and want to process several samples at once. You are most comfortable basing titers on viral genome content.

More Information

Applications

  • Determination of baculovirus titers to optimize recombinant protein expression
  • AAV production
  • Protein vaccine production

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 Analysis and Kit Components Lists are located under the Documents tab.


BacPAK p35 ELISA Kit

Learn how to obtain a functional baculovirus titer in <20 hr with the BacPAK p35 ELISA Kit


BacPAK Baculovirus GoStix Plus

Watch the BacPAK Baculovirus GoStix Plus in action to easily titer your baculovirus


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BacPAK p35 ELISA Kit

Learn how to obtain a functional baculovirus titer in <20 hr with the BacPAK p35 ELISA Kit

BacPAK Baculovirus GoStix Plus

Watch the BacPAK Baculovirus GoStix Plus in action to easily titer your baculovirus

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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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