Anti-Baculovirus Envelope gp-64 from eBioscience

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review
Structural Biology
Stanford University
Post-Doc
Fluorescence Activated Cell Sorting (FACS) plot showing data from four insect cell cultures. Shown are results from a high baculoviral titre supernatant (green) and a low baculoviral supernatant (orange). Positive (light blue) and negative (dark blue) controls are included for comparison.

Company:

eBioscience

Product Name:

Anti-Baculovirus Envelope gp-64

Catalog Number:

12-6991-80

Image

This antibody is used for the detection of the baculovirus envelope protein gp-64. GP-64 protein is upregulated on the surface of insect cells shortly after transfection with baculovirus. Insect cells are widely used for the production of recombinant mammalian proteins and this protein may be used as a marker of successful transfection without the need for full-scale protein preparation.

Experimental Design and Results Summary

Applications

Detection via flow cytometry of gp-64 expressed at the cell surface

Sample

Hi5 insect cells, transfected with high-titer baculoviral supernatant

Primary Incubation

Seed one well (for each transfectant to be tested) with 2 x 10^6 Hi5 insect cells. Add 100ul of the baculoviral supernatant to be tested to each well and shake, covered and sealed for 24 hours at 27°C. Spin down and wash cells from each well twice. Resuspend in approx. 25ul of FACS buffer and add 0.5ul (0.1ug) anti-gp64 antibody to the cells and incubate at 4°C for 30min. Wash cells prior to flow cytometry analysis for staining.

Blocking Agent

None

Secondary Incubation

N/A

Tertiary Incubation

N/A

Detection

PE-conjugated

Results Summary

This antibody can be used as a successful surrogate for recombinant protein production in an insect cell system. The pattern of staining allows the used to distinguish supernatant with high, medium and low baculoviral titre without the need for quantification of protein production.

Additional Notes

None

Image Gallery

Fluorescence Activated Cell Sorting (FACS) plot showing data from four insect cell cultures. Shown are results from a high baculoviral titre supernatant (green) and a low baculoviral supernatant (orange). Positive (light blue) and negative (dark blue) controls are included for comparison.

Summary

The Good

A reliable and simple antibody that can save time and money in insect cell protein preps.

The Bad

None.

The Bottom Line

A useful adjunct for labs producing recombinant proteins in insect cells.

Share your experience with other scientists. Write a Review! »

Join the discussion