Anti nlrp3 antibody

Geniposide (lot number: 110749-201718), Ginsenoside Rg1 (lot number:110703-201832), Ginsenoside Re (lot number:1110754-201626), Notoginsenoside R1 (lot number: 110745-201619) were purchased from the National Institutes for Food and Drug Control (Beijing, China).
NLRP3 inflammasome inhibitor (MCC950) and 2% 2,3,5-triphenyltetrazolium chloride (TTC) solution were purchased from Sigma-Aldrich (USA). The primary antibodies were provided as follows: mouse monoclonal anti-Bad antibody (sc-8044), mouse monoclonal anti-ASC antibody (sc-271054) were purchased from Santa Cruz Biotechnology (USA). Rat monoclonal anti-BcL-XL (2764), anti-Caspase 1 (4199) were purchased from Cell Signaling Technology (USA). Rabbit polyclonal anti-IL-1β antibody (ab9722), rabbit polyclonal anti-IL-18 antibody (ab71495) and mouse monoclonal anti-NeuN antibody (ab104224) were purchased from Abcam (USA). Rabbit polyclonal Anti-NLRP3 antibody was purchased from Novus (USA). GAPDH (YM3029) was purchased from Immunoway (USA).

Immunohistochemistry for kidney was carried out as previously reported.^{54 (link), 55 (link)} Simply, the left kidney was cut in half transversely, fixed in 10% formalin and embedded in paraffin. The kidney sections (5 μm) were deparaffinized, rehydrated, blocked with 3% BSA and incubated with anti-BCL6 antibody (GeneTex Inc., Irvine, CA, USA) or anti-NLRP3 antibody (Novus Biologicals, Littleton, CO, USA). Then biotinylated secondary antibodies were used and followed by 3,3’-diaminobenzidine (DAB) solution to detect the avidin–biotin complex signal. Counterstaining was then performed before examination under a light microscope (DP70, Olympus, Tokyo, Japan).

After the carotid arteries were dried and dewaxed using dimethyl benzene, arteries were rehydrated with graded ethanol solutions. The endogenous peroxidase activity was blocked using 3% hydrogen peroxide for 20 min. After treatment with fetal bovine serum, the arteries were incubated overnight at 4°C with anti-NLRP3 antibody (Novus, Littleton, Co, USA) and anti-p62 antibody (Abcam, Cambridge, MA, USA). The samples were then incubated with a secondary antibody for 1 h at room temperature.