Human Caspase-9 Antibody

Capture of Human Caspase-9 and Human Caspase-9 complexed with APAF-1 detected by Western Blot. Western blot shows Jurkat human acute T cell leukemia cell line lysates untreated (-) or treated (+) with 50 mM dATP and 1 mg/mL rat cytochrome c for 60 minutes, then captured on a 6-well dish coated at 10 µg/mL with Mouse Anti-Human Caspase-9 Monoclonal Antibody (Catalog # MAB8301). PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Caspase-9 Monoclonal Antibody (Catalog # MAB8301, left side) or Mouse Anti-Human APAF-1 Monoclonal Antibody (MAB868, right side) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF007). Specific bands were detected for Caspase-9 Precursor at approximately 46 kDa and the Caspase-9 p37 subunit at approximately 37 kDa (as indicated). A specific band was detected for APAF-1, captured as part of Caspase-9 complexed with APAF-1, at approximately 135 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.

Detection of Human Caspase‑9 by Simple Western^TM. Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 mM Staurosporine (STS) for 3 hours, loaded at 0.2 mg/mL. A specific band was detected for Caspase‑9 at approximately 53 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human Caspase‑9 Monoclonal Antibody (Catalog # MAB8301). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

Detection of Human Caspase‑9 by Western Blot. Western blot shows lysates of HEK293T human embryonic kidney cell line and HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Caspase‑9 Monoclonal Antibody (Catalog # MAB8301) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Caspase‑9 at approximately 46 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 4.

Detection of Human Caspase‑9 by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 ug/ml Staurosporine (STS) for 2 hours. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Caspase‑9 Monoclonal Antibody (Catalog # MAB8301) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Caspase‑9 at approximately 46, 37, 35 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 4.

Detection of Human Caspase-9 by Western Blot Sorbitol-dependent TDP-43 relocalization is reversible and not dependent on Nup153 cleavage.(A) Western blot of nuclear pore complex protein Nup153, nuclear lamina component lamin B1, procaspases -3 and -9 and their activated forms caspase-3 and caspase-9 in HeLa cells treated with sorbitol after pre-treatment with DMSO control or caspase inhibitor Z-VAD-FMK. alpha -tubulin was used as a loading control. (B) Immunofluorescence staining of nuclear-cytoplasmic shuttling proteins TDP-43 (green) and HuR (red) in sorbitol-stressed HeLa cells after 0 min, 15 min or 60 min rescue in normal medium. Scale bar = 30 μm. (C) Quantification of cytoplasmic TDP-43 and HuR protein in sorbitol-treated HeLa cells after 0 min, 15 min or 60 min rescue (N = 3). Results represent mean percentage of cytoplasmic signal ± SEM;** p < 0.01, *** p < 0.001, **** p < 0.0001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28510586), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Human Caspase-9 Antibody by Western Blot Sorbitol-dependent TDP-43 relocalization is reversible and not dependent on Nup153 cleavage.(A) Western blot of nuclear pore complex protein Nup153, nuclear lamina component lamin B1, procaspases -3 and -9 and their activated forms caspase-3 and caspase-9 in HeLa cells treated with sorbitol after pre-treatment with DMSO control or caspase inhibitor Z-VAD-FMK. alpha -tubulin was used as a loading control. (B) Immunofluorescence staining of nuclear-cytoplasmic shuttling proteins TDP-43 (green) and HuR (red) in sorbitol-stressed HeLa cells after 0 min, 15 min or 60 min rescue in normal medium. Scale bar = 30 μm. (C) Quantification of cytoplasmic TDP-43 and HuR protein in sorbitol-treated HeLa cells after 0 min, 15 min or 60 min rescue (N = 3). Results represent mean percentage of cytoplasmic signal ± SEM;** p < 0.01, *** p < 0.001, **** p < 0.0001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28510586), licensed under a CC-BY license. Not internally tested by R&D Systems.