Human/Mouse PARP Antibody

Detection of Human PARP by Western Blot.

Western blot shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated with anti-Fas and 1 µM staurosporine (STS) for 0 to 4 hours (h; as indicated). PVDF membrane was probed with 0.6 µg/mL of Rat Anti-Human/Mouse PARP Monoclonal Antibody (Catalog # MAB600) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). Specific bands were detected for PARP at approximately 116 kDa and the 84 kDa product of PARP cleavage (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Detection of Mouse PARP by Western Blot.

Western blot shows lysates of NIH-3T3 mouse embryonic fibroblast cell line treated with 1 µM staurosporine (STS) for 0 to 4 hours (h; as indicated). PVDF membrane was probed with 0.6 µg/mL of Rat Anti-Human/Mouse PARP Monoclonal Antibody (Catalog # MAB600) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). Specific bands were detected for PARP at approximately 116 kDa and the 60-70 kDa bands generated with staurosporine treatment (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Detection of Human PARP by Western Blot

The treatment of n-BP rescued the progression of QA-induced excitotoxicity in SCA3 PPs. The control and/or SCA3 PPs were treated with or without QA (1 μM) in the presence of n-BP (0, 10, 20, and 40 μg/mL) for 12 h. (A) Images of QA-treated SCA3 PPs in the presence of BP (40 μg/mL). (B) Confocal images of poly Q localization in BP (40 μg/mL)-treated SCA3 PPs in the presence of QA (1 um). When compared with Figure 2C, 40 μg/mL of n-BP prevented the amounts of colocalized polyQ in the cell nucleus of PPs. The distribution of polyQ in the nucleus is indicated by white arrowheads. Scale bar = 100 μm; (C) protein analysis of wild-type, mutant ATXN3 and its proteolytic fragment in cell lysates of control and SCA3 PPs, as assessed by immunoblots. Soluble, soluble protein; Insoluble, insoluble protein; (D,E) representative immunoblot of control and cleaved PARP1 in cell lysates of control and SCA3 PPs, as assessed by immunoblots. The HDAC2 were used as loading controls. The quantification from three independent images is presented as the means ± standard deviation. t(4) = 2.824, p < 0.05, for lane 1 vs. lane 2; t(4) = 5.613, p < 0.01, for lane 1 vs. lane 3; t(4) = 3.522, p < 0.05, for lane 1 vs. lane 4; (F) the quantification result of calcium concentration in SCA3 PPs, as assessed by Fura-2 indicator. Cells were treated with QA (1 μM) in the presence of n-BP (0, 10, 20, and 40 μg/mL). Data are presented as the means ± standard deviation. t(4) = 2.95, p < 0.05; (G) the calpain activity in control and SCA3 PPs cell lysates, as assessed by ELISA assay. The quantification results from three independent replicates are presented as the means ± standard deviation. t(4) = 3.27, p < 0.05, for lane 3 vs. lane 4; t(4) = 17.55, p < 0.01, for lane 3 vs. lane 5; t(4) = 9.019, p < 0.01, for lane 3 vs. lane 6. (H) Protein analysis of calpain 1, calpain 2 and calpastatin in cell lysates of SCA3 PPs, as assessed by immunoblots. Cells were treated with or without QA (1 μM) in the presence of n-BP (0, 10, 20, and 40 μg/mL). *, p < 0.05; **, p < 0.01. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35163312), licensed under a CC-BY license. Not internally tested by R&D Systems.