Mouse monoclonal anti parp

Western blot analysis of whole cell lysate was performed as reported in [41 (link)]. The following primary antibodies were employed in the study: rabbit monoclonal anti-NF-κB/RelA and rabbit monoclonal anti-LC3 (both from Cell Signaling Technology, Beverly, MA, USA); mouse monoclonal anti-β-actin (Sigma-Aldrich); rabbit polyclonal anti-phospho-NF-κB/p65 (S529, Abcam, Cambridge, MA, USA); mouse monoclonal anti-PARP and mouse monoclonal anti-HIF-1α (both from BD Biosciences, San Jose, CA, USA).

Western blot analysis was performed as previously reported [26 (link)]. In brief, total cellular protein concentration was determined using the BCA method then separated by SDS-PAGE and transferred to nitrocellulose (Schleicher and Schuell, Keene, NH). The primary antibodies used were rabbit monoclonal anti-XIAP, rabbit monoclonal anti-Smac/DIABLO, rabbit monoclonal anti-cytochrome c, rabbit monoclonal anti-c-IAP1, rabbit monoclonal anti-activated caspase-3, mouse monoclonal anti-PARP (BD Pharmingen, USA), rabbit monoclonal anti-AIF, rabbit monoclonal anti-pSmad1/5, (Cell signaling Technology, MA, USA), rabbit monoclonal anti-Id1 (Calbioreagents, San Mateo, CA), rabbit polyclonal anti-Smad 1/5 (Upstate Biotechnology, NY, USA), rabbit polyclonal survivin (Novus Biologicals, CO, USA), rabbit anti-actin, an affinity isolated antigen specific antibody (Sigma, Saint Louis, MO), rabbit polyclonal anti-GAPDH (Sigma) and mouse monoclonal anti-Spectrin (EMD Millipore, CA, USA).
Blots suggesting regulation of loading controls actin or GAPDH by Ym155 and/or JL5 were then probed with spectrin, which is also a cytoskeletal protein.

Mouse monoclonal anti-PARP (1:500), anti-beclin-1 (1:500), rabbit polyclonal anti-Bax (1:2000), and anti-Bak (1:3000) were from BD Biosciences (San Jose, CA, USA); mouse monoclonal anti-Bcl-xL (1:1000), rabbit polyclonal anti-Mcl-1 (1:1000), anti-cyclin B1 (1:500), and anti-p-histone H3 (Ser10) (1:1000) were from Santa Cruz (Santa Cruz, CA, USA); mouse monoclonal anti-β-actin (1:10,000), rabbit polyclonal anti-LC3B (1:2000), and anti-p62 (1:2000) were from Sigma; rabbit polyclonal anti-cleaved caspase-9 (Asp315) (1:500) and anti-cleaved caspase-3 (Asp175) (1:500) were from Cell Signaling (Danvers, MA, USA).