Baculovirus expression system: a complete system

Substrate for β-glucuronidase, an acid hydrolase encoded by the gusA gene. When used in histochemical assay methods, X-GLUC will give a blue precipitate in the presence of β-glucuronidase.

Required Products

Cat. #	Product	Size	Price	License	Quantity	Details
631411	IPLB-Sf21 Insect Cells	1 Vial	USD $395.00
IPLB-Sf21 insect cells may be used as a host for propagating the Autographa californica multiple-enveloped nuclear polyhedrosis virus (AcMNPV) and its expression vector derivatives generated from ourBacPAK system. The IPLB-Sf21 cell line isderived from pupal ovaries of the fall armyworm, Spodoptera frugiperda. Exponentially growing IPLB-Sf21 cells are concentrated by centrifugation and frozen in insect cell complete medium containing 10% dimethysulfoxide (DMSO). Documents Components Image Data Back 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). 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). 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

Complete kit for expressing recombinant proteins at high levels. The BacPAK System uses the baculovirus Autographa californica nuclear polyhedrosis virus (AcMNPV) to produce target proteins in insect cells. More than 90% of the viruses produced by the transfected cells carry the target protein. The expressed recombinant protein is similar in structure, biological activity, and immunological reactivity to the naturally occurring protein because insect host cells provide post-translational processing similar to that of mammalian cells.

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BacPAK6 Viral DNA map

BacPAK6 Viral DNA map.

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Protein production from recombinant viruses generated using the BacPAK Baculovirus Expression System

Protein production from recombinant viruses generated using the BacPAK Baculovirus Expression System. Recombinant viruses were obtained by cotransfection of transfer vectors with BacPAK6 Viral DNA (Bsu36 I digest), followed by amplification in Sf21 cells. The SDS PAGE analysis of cellular lysates was performed 48 hr after infection of the Sf21 cultures. Lane 1: uninfected Sf21 cells. Lane 2: Sf21 cells infected with wild-type AcMNPV virus. Lane 3: Sf21 cells infected with nonrecombinant BacPAK6 virus. Lane 4: Sf21 cells infected with BacPAK8-GUS recombinant virus. Lane 5: purified CAT protein. Lane 6: Sf21 cells infected with BacPAK9-CAT recombinant virus. Lane M: molecular weight marker.

Required Products

Cat. #	Product	Size	Price	License	Quantity	Details
631411	IPLB-Sf21 Insect Cells	1 Vial	USD $395.00
IPLB-Sf21 insect cells may be used as a host for propagating the Autographa californica multiple-enveloped nuclear polyhedrosis virus (AcMNPV) and its expression vector derivatives generated from ourBacPAK system. The IPLB-Sf21 cell line isderived from pupal ovaries of the fall armyworm, Spodoptera frugiperda. Exponentially growing IPLB-Sf21 cells are concentrated by centrifugation and frozen in insect cell complete medium containing 10% dimethysulfoxide (DMSO). Documents Components Image Data Back 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). 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). 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

Baculovirus DNA specially designed and prepared to give a high proportion of recombinant viral expression vectors. The Bsu36 I-digested BacPAK6 DNA is ready-to-use and is sufficient for 5 transfections. The transfection reagent Bacfectin is also provided.

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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. 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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631401: BacPAK6 DNA (Bsu36 I digest)

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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). 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). 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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BacPAK6 Viral DNA map

BacPAK6 Viral DNA map.

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Protein production from recombinant viruses generated using the BacPAK Baculovirus Expression System

Protein production from recombinant viruses generated using the BacPAK Baculovirus Expression System. Recombinant viruses were obtained by cotransfection of transfer vectors with BacPAK6 Viral DNA (Bsu36 I digest), followed by amplification in Sf21 cells. The SDS PAGE analysis of cellular lysates was performed 48 hr after infection of the Sf21 cultures. Lane 1: uninfected Sf21 cells. Lane 2: Sf21 cells infected with wild-type AcMNPV virus. Lane 3: Sf21 cells infected with nonrecombinant BacPAK6 virus. Lane 4: Sf21 cells infected with BacPAK8-GUS recombinant virus. Lane 5: purified CAT protein. Lane 6: Sf21 cells infected with BacPAK9-CAT recombinant virus. Lane M: molecular weight marker.

Required Products

Cat. #	Product	Size	Price	License	Quantity	Details
631411	IPLB-Sf21 Insect Cells	1 Vial	USD $395.00
IPLB-Sf21 insect cells may be used as a host for propagating the Autographa californica multiple-enveloped nuclear polyhedrosis virus (AcMNPV) and its expression vector derivatives generated from ourBacPAK system. The IPLB-Sf21 cell line isderived from pupal ovaries of the fall armyworm, Spodoptera frugiperda. Exponentially growing IPLB-Sf21 cells are concentrated by centrifugation and frozen in insect cell complete medium containing 10% dimethysulfoxide (DMSO). Documents Components Image Data Back 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). 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). 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

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.