Fas Ligand Antibody from GeneTex

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Microbiology-Immunology
Northwestern University
PhD Student
FasL (0.2 ug) is readily detected using this antibody (1:1000) by Western blot between 30-35 kDa due to varying degrees of glycosylation. Dimerized protein can also be observed at 70 kDa.

Company:

GeneTex

Product Name:

Fas Ligand Antibody (N2C1)

Catalog Number:

GTX100644

Image

This antibody was used to test Fas Ligand (FasL) stability during various treatments as assessed by Western blot. We initially screened multiple antibodies from various vendors and this product worked the best.

Experimental Design and Results Summary

Applications

Western blotting

Sample

Human recombinant FasL (aa 103-281) produced in HEK 293T cells.

Primary Incubation

Fas Ligand antibody (N2C1) supplied by GeneTex. Antibody was diluted 1:1000 in blocking agent and incubated overnight at 4°C.

Blocking Agent

5% nonfat milk in TBST, incubated 30 minutes at room temperature.

Secondary Incubation

Goat anti-rabbit IgG-HRP supplied by Sigma-Aldrich(catalog #A6154). Antibody was diluted 1:2000 in blocking agent and incubated for 1 hour at room temperature.

Tertiary Incubation

N/A

Detection

ECL Prime Western Blotting Detection Reagent (GE Amersham, #RPN2232) used following manufacturer instructions.

Results Summary

No background or non-specific bands are detected and signal intensity is generally strong. Because this antibody is polyclonal, FasL fragments may also be detected.

Additional Notes

0.2 ug protein can generally be detected on film using exposures of less than 1 minute. FasL is modified through the addition of 3 glycosylations, which result in full-length bands between 30-35 kDa.

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FasL (0.2 ug) is readily detected using this antibody (1:1000) by Western blot between 30-35 kDa due to varying degrees of glycosylation. Dimerized protein can also be observed at 70 kDa.

Summary

The Good

Great signal with no non-specific bands.

The Bad

None.

The Bottom Line

Works as expected.

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