This is a knockout-validated antibody summary, based on publications "Combination of IAP antagonist and IFNγ activates novel caspase-10- and RIPK1-dependent cell death pathways", as cited below [1], and "Microbial signals, MyD88, and lymphotoxin drive TNF-independent intestinal epithelial tissue damage" for western blot knockout validation (figure 6d) [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: Cleaved Caspase-8 (Asp387)

Catalog number: 8592

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues surrounding Asp387 of mouse caspase-8 protein. Reacts with endogenous levels of mouse caspase-8 only when cleaved at Asp387. In addition to the p18 subunit, this antibody also detects cleavage products containing the pro-domain with the p18 subunit (p43). Suitable for western blot, immunoprecipitation, immunofluorescence/immunocytochemistry, and flow cytometry.

Western blot

WT and Casp8−/− mouse dermal fibroblasts (MDFs). Cells were lysed in DISC lysis buffer on ice for 20 min before the addition of SDS sample loading buffer or lysed directly in SDS lysis buffer (126 mM Tris-HCl, pH 8, 20% (v/v) glycerol, 4% (w/v) SDS, 0.02% (w/v) bromophenol blue, 5% (v/v) 2-mercaptoethanol) boiled and sonicated.

5% milk in PBST.

Diluted in 2% BSA in PBST.

Wild-type and Casp8−/− MDFs after the indicated treatments were analysed by immunoblotting (n=2). Arrows indicate full-length and processing products of caspases. Please see Figure 3d in the article [1].