Human/Mouse/Rat STAT5b Antibody

Detection of Human, Mouse, and Rat STAT5b by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, Daudi human Burkitt's lymphoma cell line, NIH-3T3 mouse embryonic fibroblast cell line, M1 mouse myeloid leukemia cell line, and PC-12 rat adrenal pheochromocytoma cell line. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse/Rat STAT5b Monoclonal Antibody (Catalog # MAB15841) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for STAT5b at approximately 90 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

STAT5b in HepG2 Human Cell Line. STAT5b was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Rabbit Anti-Human/Mouse/Rat STAT5b Monoclonal Antibody (Catalog # MAB15841) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human and Mouse STAT5b by Simple Western^TM. Simple Western lane view shows lysates of M1 mouse myeloid leukemia cell line and Daudi human Burkitt's lymphoma cell line, loaded at 0.2 mg/mL. A specific band was detected for STAT5b at approximately 90 kDa (as indicated) using 1 μg/mL of Rabbit Anti-Human/Mouse/Rat STAT5b Monoclonal Antibody (Catalog # MAB15841). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

STAT5b Specificity is Shown by Immunocytochemistry in Knockout Cell Line. STAT5b was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with IFN-alpha 1 but is not detected in STAT5b knockout (KO) HeLa cell line using Rabbit Anti-Human/Mouse/Rat STAT5b Monoclonal Antibody (Catalog # MAB15841) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Western Blot Shows Human STAT5b Specificity by Using Knockout Cell Lines. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line, STAT1 knockout (KO) HeLa cell line, STAT2 KO HeLa cell line, STAT3 KO HeLa cell line, STAT5a KO HeLa cell line, STAT5b KO HeLa cell line, and STAT6 KO HeLa cell line. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse/Rat STAT5b Monoclonal Antibody (Catalog # MAB15841) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for STAT5b at approximately 85 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the STAT5b knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Western Blot Shows Human STAT5b Specificity Using Knockout Cell Line. Western blot shows lysates of HAP1 human near-haploid cell line and STAT5b knockout HAP1 cell line (KO). Nitrocellulose membrane was probed with 1 µg/mL of Rabbit Anti-Human/Mouse/Rat STAT5b Monoclonal Antibody (Catalog # MAB15841) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody. A specific band was detected for STAT5b at approximately 85 kDa (as indicated) in the parental HAP1 cell line, but is not detectable in knockout HAP1 cell line. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.

Detection of Stat5b by Immunoprecipitation. Immunoprecipitation was performed on cell lysate of HAP1 human near-haploid cell line using 2.0 μg of Rabbit Anti-Human Stat5b Monoclonal Antibody (Catalog # MAB15841) pre-coupled to protein G or protein A beads. Immunoprecipitated Stat5b was detected with a Mouse Anti-Stat5b antibody. The Ponceau stained transfers of each blot are shown. SM=10% starting material; UB=10% unbound fraction; IP=immunoprecipitated. Image, protocol, and testing courtesy of YCharOS Inc. (ycharos.com).