Horseradish peroxidase conjugated goat anti rabbit sc 2004

NLRP3 (AG-20B-0014, AdipoGen, CA, USA); IL1β (ab9722, Abcam, MA, USA); Anti-CD63 antibody (ab216130, Abcam, MA, USA); Anti-CD9 antibody (ab92726, Abcam, MA, USA), Anti-TSG101 antibody (ab125011, Abcam, MA, USA), Anti-Alix antibody (ab275377, Abcam, MA, USA), Anti-Calnexin antibody (ab133615, Abcam, MA, USA); Anti-PSD95 antibody (ab2723, Abcam, MA, USA); Anti-GAD65 antibody (ab239372, Abcam, MA, USA); Anti-Gephyrin antibody (ab181382, Abcam, MA, USA); Anti-vGLUT1 antibody (AB5905, Millipore, Burlington, MA, USA), GFP expressing plasmid (13031, Adgene, Watertown, MA, USA); β-actin (A5316, Sigma- Aldrich, MO, USA); horseradish peroxidase conjugated goat anti-rabbit (sc-2004, Santa Cruz Biotechnology, TX, USA) and horseradish peroxidase conjugated goat anti-mouse (sc-2005, Santa Cruz Biotechnology, TX, USA); human primary microglia (Cat # 1900; Sciencell research laboratory, CA, USA) and BV2 microglial cell line was received from Dr. Sanjay Maggirwar (University of Rochester Medical Center, Rochester, NY, USA).

Protein expression analysis was detected by Western blot analysis as previously described [29 (link), 40 (link)]. The membranes were incubated with primary antibodies, including anti-BMF (ab9655, 1:1000, Abcam), anti-Runx2 (ab76956, 1:1000, Abcam), anti-p16 (10883-1-AP, 1:1000, Proteintech), anti-p21 (10355-1-AP, 1:1000, Proteintech), and anti-GAPDH (sc-365062, 1:3000, Santa Cruz Biotechnology) at 4°C overnight, followed by incubation with the horseradish peroxidase-conjugated goat anti-rabbit (sc-2004, 1:5000, Santa Cruz Biotechnology) or goat anti-mouse (sc-2005, 1:5000, Santa Cruz Biotechnology) secondary antibodies at 37°C for 1 hour. The immunoreactive bands were visualized using the ECL Plus Western blot detection kit (Amersham Biosciences U.K. Ltd) and densitometric quantification of band intensity from three independent experiments was carried out with the Image-Pro Plus 6.0 software. The relative expression level of target protein was normalized to the intensity of the GAPDH band.