| 货号 | AF1584 |
| 别名 | signal transducer and activator of transcription 5B; STAT5; transcription factor STAT5B | 全称 | Signal Transducer and Activator of Transcription 5b |
| 反应种属 | Human/Mouse |
| 应用 | Western Blot,Simple Western,Immunoprecipitation,CyTOF-ready,Immunocytochemistry,Intracellular Staining by Flow Cytometry |
| 目标/特异性 | Detects human and mouse STAT5b. |
| 使用方法 | Western Blot: 0.2 µg/mL Simple Western: 10 µg/mL Immunoprecipitation: 10 µg/106cells Immunocytochemistry: 5-15 µg/mL Intracellular Staining by Flow Cytometry: 2.5 µg/106cells |
| 来源 | Reconstitute at 0.2 mg/mL in sterile PBS. |
| 产品组分 |
| 供应商 | R&D Systems |
| Entrez Gene IDs | 6777 (Human); 20851 (Mouse) |
| 应用文献 | |
| R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions. Rac1 and a GTPase-activating protein, MgcRacGAP, are required for nuclear translocation of STAT transcription factors. | |
| 纯化方式 | Antigen Affinity-purified |
| 免疫原 | Human STAT5b synthetic peptide aa 777-787 |
| 生物活性 | Human, Mouse |
| 标记 | Unconjugated |
| 溶解方法 | Reconstitute at 0.2 mg/mL in sterile PBS. |
| 背景 | Signal transduction and activator of transcription 5 (STAT5) is a member of the Jak/STAT signal transduction pathway and is activated by a variety of cytokines (IL22, IL6, IFN-α). STAT5 has two isoforms (A and B) that share 93% amino acid identity and bind the DNA consensus site TTCN3GAA. STAT5 mediates cytokine signaling by acting as a signal transducer in the cytoplasm and, upon phosphorylation, translocates to the nucleus and activates transcription of specific genes. STAT5 is involved in a wide array of biological processes ranging from regulating apoptosis to adult mammary gland proliferation, differentiation and survival. |
| 运输条件 | Blue Ice |
| 存放说明 | -20℃ |
| 参考文献 |
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Detection of Human, Mouse, and Rat STAT5b by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, Daudi human Burkitts 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 0.2 µg/mL of Rabbit Anti-Human/Mouse STAT5b Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1584) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # 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 STAT5b Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1584) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI(blue). Specific staining was localized to cytoplasm and nucleus. View our protocol for Fluorescent ICC Staining of Cells on Coverslips. | |
Immunoprecipitation of Human and Mouse STAT5b. HeLa human cervical epithelial carcinoma and M1 mouse myeloid leukemia cell line were untreated (-) or activated (+) with 100 ng/mL Recombinant Human IFN‑ gamma (Catalog # 285-IF) for 15 minutes. STAT5b was immunoprecipitated from lysates of 5 x 106 cells following incubation with 2 µg Rabbit Anti-Human/Mouse STAT5b Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1584) for 1 hour at room temperature. STAT5b-antibody complexes were absorbed using Protein A sepharose (Invitrogen, Catalog # 10-1041). Immunoprecipitated STAT5b was detected by Western blot using 1 µg/mL Human Phospho-STAT5a/b (Y699) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4190). View our recommended buffer recipes for immunoprecipitation. | |
Detection of STAT5b in Jurkat Human Cell Line by Flow Cytometry. Jurkat human acute T cell leukemia cell line was stained with Rabbit Anti-Human/Mouse STAT5b Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF1584, filled histogram) or control antibody (Catalog # AB-105-C, open histogram), followed by Allophycocyanin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0111). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol. | |
Detection of Human and Mouse STAT5b by Simple WesternTM. Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and NIH‑3T3 mouse embryonic fibroblast cell line, loaded at 0.2 mg/mL. A specific band was detected for STAT5b at approximately 92 kDa (as indicated) using 10 µg/mL of Rabbit Anti-Human/Mouse STAT5b Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1584). 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. |
