This is a knockout-validated antibody summary, based on the publications "Familial Mediterranean fever mutations lift the obligatory requirement for microtubules in Pyrin inflammasome activation", as cited below [1], "Inflammasome Is Activated in the Liver of Cholestatic Patients and Aggravates Hepatic Injury in Bile Duct-Ligated Mouse" for western blot knockout validation (figure 2c) [2], "Interferon-γ primes macrophages for pathogen ligand-induced killing via a caspase-8 and mitochondrial cell death pathway" for western blot knockout validation (figure s6c) [3], and and "CrkII/Abl phosphorylation cascade is critical for NLRC4 inflammasome activity and is blocked by Pseudomonas aeruginosa ExoT" for western blot knockout validation (figure s7a) [4]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Mouse monoclonal IgG1

Company: Adipogen

Antibody: Caspase-1 (p20)

Catalog number: AG-20B-0042-C100

Summary: Mouse monoclonal IgG1 against recombinant mouse caspase-1. Reacts with mouse by western blot, immunoprecipitation and immunohistochemistry.

Western blot

Wild-type and Casp1−/− bone marrow-derived macrophages (BMDMs). Cell lysates and culture supernatants were incubated with cell lysis buffer [5].

0.05% Tween-20 (vol/vol) and 3% (wt/vol) nonfat dry milk.

1:1,000 dilution in blocking buffer overnight.

HRP-conjugated secondary anti-mouse or anti-rabbit antibodies (111-035-144 and 112-035-143; 1:5,000, Jackson ImmunoResearch Laboratories)

Enhanced chemiluminescence (Thermo Scientific).

Combined supernatant and cell lysates from wild-type and Casp1−/− BMDMs were treated as indicated and immunoblotted for caspase-1 with β-actin as loading control. Please see Figure S2c in the article [1].