Santa Cruz Biotechnology
FADD Antibody (H-181)
sc-5559
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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.
Western blotting
Jurkat whole cell lysate
FADD antibody supplied by Santa Cruz Biotech (catalog #sc-5559). Antibody was diluted 1:1000 in blocking agent and incubated overnight at 4°C.
5% nonfat milk in Tris-Buffered Saline with Tween (TBST), incubated 30 minutes at room temperature.
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.
N/A
ECL Prime Western Blotting Detection Reagent (GE Amersham, cat #RPN2232) used following manufacturer instructions.
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.
Jurkat cells were lysed with RIPA buffer.
Strong signal.
Some non-specific binding.
Works as expected.