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.

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.

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.

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

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

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