Cells were dissolved on ice in 250 µL ice‐cold lysis buffer (CelLytic M; Sigma‐Aldrich) containing protease inhibitor (Sigma‐Aldrich) as described previously.19 , 21 Equal amounts of protein (50 µg/well) were separated on a discontinuous SDS‐10% polyacrylamide gel and blotted onto a nitrocellulose membrane (Bio‐Rad). Western blot analysis was carried out using the following primary Abs: G‐9 (Santa Cruz Biotechnology) for Fas; FD19 (Santa Cruz Biotechnology) and anti‐FADD rabbit Ab (Cell Signaling Technology) for FADD; 1C12 (Cell Signaling Technology) and anti‐caspase‐8 rabbit Ab (GeneTex) for procaspase‐8/FLICE and caspase‐8; 5D8 (Santa Cruz Biotechnology) for c‐FLIPL; anti‐c‐FLIPγ/δ rabbit Ab (Sigma‐Aldrich) for c‐FLIPS; D5J1E (Cell Signaling Technology) for both forms and the cleaved c‐FLIPL; 38 (BD Bioscience) for RIP1; C‐2 (Santa Cruz Biotechnology) for BCL2; E18 (Abcam) for BCL‐xL; Y37 (Abcam) for MCL1; 31A1067 (Santa Cruz Biotechnology) for procaspase‐3 and the cleaved forms; 194C1439 (Santa Cruz Biotechnology) for cleaved PARP1; and AC74 (Sigma‐Aldrich) for β‐actin. Antibody signals were visualized using Western Blue (Promega). We quantified the expression levels by comparison with each β‐actin level using ImageJ version 1.52k (://imagej.nih.gov/ij/ ).
Anti fadd rabbit ab
Manufactured by Cell Signaling Technology
Anti-FADD rabbit Ab is a primary antibody that recognizes the FADD (Fas-Associated protein with Death Domain) protein. FADD is an important adaptor protein involved in various cell signaling pathways. This antibody can be used to detect and study the FADD protein in various experimental applications.
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2 protocols using anti fadd rabbit ab
1
Western Blot Analysis of Apoptosis Regulators
2
Comprehensive Fas/FasL Signaling Antibody Panel
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We used the following primary Abs: anti‐Fas mouse (Ms) IgM 7C11 (Beckman Coulter) or CH‐11 (MBL); anti‐Fas Ms IgG1 ZB4 (Beckman Coulter); anti‐Fas Ms IgG1 UB2 (Beckman Coulter); anti‐Fas Ms IgG1 G‐9 (Santa Cruz Biotechnology); anti‐Fas rabbit Ab C‐20 (Santa Cruz Biotechnology); anti‐FasL Ms IgG1 NOK‐1 (BD Biosciences); anti‐FasL rabbit Ab Q‐20 (Santa Cruz Biotechnology); anti‐FADD Ms IgG1 FD19 (Santa Cruz Biotechnology); anti‐FADD rabbit Ab (Cell Signaling Technology); anti‐caspase‐8 Ms IgG1 1C12 (Cell Signaling Technology); anti‐caspase‐8 rabbit IgG (GeneTex); anti‐c‐FLIPL Ms IgG3 5D8 (Santa Cruz Biotechnology); anti‐c‐FLIPγ/δ rabbit Ab (Sigma‐Aldrich); anti‐c‐FLIP rabbit IgG D5J1E (Cell Signaling Technology); anti‐c‐FLIP Ms IgG1 NF6 (AdipoGen); anti‐RIP Ms IgG2a 38 (BD Bioscience); anti‐BCL2 Ms IgG1 C‐2 (Santa Cruz Biotechnology); anti‐BCL‐xL rabbit IgG E18 (Abcam); anti‐MCL1 rabbit IgG Y37 (Abcam); anti‐caspase‐3 Ms IgG1 31A1067 (Santa Cruz Biotechnology); anti‐cleaved PARP1 Ms IgG2b 194C1439 (Santa Cruz Biotechnology); and anti‐β‐actin Ms IgG2a AC74 (Sigma‐Aldrich).
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