FADD Antibody (H-181) from Santa Cruz Biotechnology

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Microbiology-Immunology
Northwestern University
PhD Student
FADD is readily detected from Jurkat cell lysates using this antibody (1:1000) by Western blotting at 27 kDa. A higher molecular weight non-specific band is also observed.

Company:

Santa Cruz Biotechnology

Product Name:

FADD Antibody (H-181)

Catalog Number:

sc-5559

Image

Our experimental goal was to determine relative levels of FADD protein production in a common laboratory T lymphocyte cell line (Jurkat). This antibody was successfully used to detect FADD by Western blot.

Experimental Design and Results Summary

Applications

Western blotting

Sample

Jurkat whole cell lysate

Primary Incubation

FADD antibody supplied by Santa Cruz Biotech (catalog #sc-5559). Antibody was diluted 1:1000 in blocking agent and incubated overnight at 4°C.

Blocking Agent

5% nonfat milk in Tris-Buffered Saline with Tween (TBST), incubated 30 minutes at room temperature.

Secondary Incubation

Goat anti-rabbit IgG-HRP (Sigma-Aldrich, cat#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, cat #RPN2232) used following manufacturer instructions.

Results Summary

FADD protein is readily detectable with a molecular weight of 27 kDa. A non-specific band of lower intensity is also observed just under 50 kDa.

Additional Notes

Jurkat cells were lysed with RIPA buffer.

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FADD is readily detected from Jurkat cell lysates using this antibody (1:1000) by Western blotting at 27 kDa. A higher molecular weight non-specific band is also observed.

Summary

The Good

Strong signal.

The Bad

Some non-specific binding.

The Bottom Line

Works as expected.

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