This is a knockout-validated antibody summary, based on the publications "Macrophage activation and polarization modify P2X7 receptor secretome influencing the inflammatory process" (Figure 1, discussed below) [1], "Macrophage activation and polarization modify P2X7 receptor secretome influencing the inflammatory process" (Figure 2) [2], "CLICs-dependent chloride efflux is an essential and proximal upstream event for NLRP3 inflammasome activation" (Figure 3) [3], "Calpain drives pyroptotic vimentin cleavage, intermediate filament loss, and cell rupture that mediates immunostimulation" for western blot knockout validation (Figure 1a) [4], and "Novel role for caspase 1 inhibitor VX765 in suppressing NLRP3 inflammasome assembly and atherosclerosis via promoting mitophagy and efferocytosis" for immunohistochemistry knockout validation (figure s5) [5]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

[enlarge]

Figure 1. Western blot of NLRP3 in bone marrow-derived macrophages (BMDMs) from wild type (WT), Casp1/11-/- or Nlrp3-/- mice primed for 4 h with LPS followed by no stimulation (−) or stimulation (+) for 20 min with ATP. PPIase levels serve as loading control. From [1].

Mouse monoclonal IgG2b

Company: Adipogen

Antibody: NLRP3/NALP3

Catalog number: AG-20B-0014

[enlarge]

Figure 2. Western blot analysis of NLRP3, caspase-1 and IL-1β proteins in total lysates (CL) and supernatants (SN) of NLRP3-/- and wild type bone-associated mesenchymal stromal cells. From figure 1g of the article [2].

Summary: Mouse monoclonal IgG2b against recombinant mouse NLRP3/NALP3 (pyrin domain/aa 1-93). Reacts with mouse and human by western blot, immunocytochemistry, immunoprecipitation, immunohistochemistry and ChIP assay.

[enlarge]

Figure 3. Immunoblot analysis of Nlrp3 and other proteins in supernatants or cell lysates of LPS-primed Nlrp3 +/+ or Nlrp3 –/– bone marrow-derived macrophages after various treatments. From [3].

Western blot

Bone marrow-derived macrophages (BMDMs) from wild type and Nlrp3-/- mice.

1:200 to 1:2,000 dilution in blocking buffer overnight at 4 C.

1:5,000 dilution HRP-conjugated secondary anti-mouse antibodies (GE Healthcare).

Pierce ECL2 kit and Thyphoon 9410 scanner.