Mouse monoclonal anti caspase 1 p20 antibody

Samples, including perihaematomal brain tissues, cell lysates and medium, were collected and subjected to western blot analysis, as described in our previous studies [16 (link), 17 (link)]. The following primary antibodies were used: rabbit polyclonal anti-CBS antibody (1:200, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-P2X7R antibody (1:1000, Alomone Labs, Jerusalem, Israel), rabbit polyclonal anti-NLRP3 antibody (1:200, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-ASC antibody (1:500, Abcam, Cambridge, MA, USA), mouse monoclonal anti-caspase-1 p20 antibody (1:200, Santa Cruz Biotechnology, CA, USA), rabbit polyclonal anti-IL-1β antibody (1:1000, Millipore, Billerica, MA, USA), rabbit polyclonal anti-myeloperoxidase antibody (anti-MPO, 1:500, Abcam, Cambridge, MA, USA) and goat polyclonal anti-Iba1 antibody (1:600, Abcam, Cambridge, MA, USA). GAPDH (1:1000, Cell Signalling Technology, Danvers, MA, USA) was used as a loading control. The proteins were detected on nitrocellulose membranes with enhanced chemiluminescence reagents (GE Healthcare, Beijing, China), and the blot bands were quantified by densitometry with Image J software (National Institutes of Health, Bethesda, MD, USA). The results are expressed as a relative density ratio, which was normalized to the mean value of the vehicle or control group.

Western blotting was performed as described previously [30 (link)]. The following primary antibodies were used: rabbit polyclonal anti-P2X7R (1:1000, Alomone Labs, Jerusalem, Israel), rabbit polyclonal anti-NLRP3 antibody (1:1000, Santa Cruz, Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-ASC antibody (1:500, Abclonal, Cambridge, MA, USA), mouse monoclonal anti-caspase-1 p20 antibody (1:1000, Santa Cruz Biotechnology), rabbit polyclonal anti-IL-1β antibody (1:1000, Millipore), rabbit monoclonal anti-IL-18 antibody (1:1000, Abcam, Cambridge, UK), mouse monoclonal anti-nitrotyrosine antibody (1:1000, Abcam), mouse monoclonal anti-iNOS antibody (1:200, Santa Cruz Biotechnology), mouse monoclonal anti-gp91^phox antibody (1:2000, BD Transduction Laboratories, San Jose, CA, USA), and rabbit polyclonal anti-myeloperoxidase antibody (MPO, 1:500, Abcam). GAPDH (1:1000, Cell Signaling Technology, Danvers, MA, USA) was employed as the loading control. Blot bands were quantified by densitometry with ImageJ software (National Institutes of Health, Baltimore, MD, USA).