This is a knockout-validated antibody summary, based on the publications "Human Cytomegalovirus Immediate Early 86-kDa Protein Blocks Transcription and Induces Degradation of the Immature Interleukin-1β Protein during Virion-Mediated Activation of the AIM2 Inflammasome", as cited below [1], and "CARD8 is an inflammasome sensor for HIV-1 protease activity" for western blot knockout validation (figure 4c) [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Immunoblot assay for NLRP3 and GAPDH in THP-1 cells transduced with Cas9 and guide RNA (gRNA) targeting HUMCYC pseudogene or in indicated CRISPR-generated knockout cells. From [1].

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: NLRP3

Catalog number: 13158

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues surrounding Pro92 of human NLRP3 protein. Reacts with human by western blot and immunoprecipitation.

Western blot

Control and NLRP3-KO THP-1 cells. Cell pellets were lysed in RIPA buffer (10 mM Tris-Cl (pH 8.0), 1 mM EDTA, 1% Triton X-100, 0.1% sodium deoxycholate, 0.1% SDS, 140 mM NaCl, and protease and phosphatase inhibitor cocktail).

5% milk in 1× phosphate-buffered saline (PBS) containing 0.1% Tween 20 for 30 min at room temperature.

Overnight at 4°C.

Horseradish peroxidase (HRP)-conjugated secondary antibodies in 5% milk for 30 min at room temperature.

Enhanced chemiluminescence (Thermo Fisher).