STAT5a/b Antibody [Unconjugated] - Pan Specific

Western blot shows lysates of K562 human chronic myelogenous leukemia cell line and M1 mouse myeloid leukemia cell line. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Human/Mouse STAT5a/b Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2168) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for STAT5a at approximately 94 kDa and STAT5b at approximately 92 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

BaF3 mouse pro-B cell line treated with 10 ng/mL Recombinant Mouse IL‑3 (Catalog # 403-ML) for 1 hour was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. STAT5a/b/DNA complexes were immunoprecipitated using 5 μg Rabbit Anti-Human/Mouse STAT5a/b Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2168) or control antibody (Catalog # AB-105-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Rabbit IgG Secondary Antibody (Catalog # BAF008). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The bcl-x promoter was detected by standard PCR.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and K562 human chronic myelogenous leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for STAT5a/b at approximately 91 kDa (as indicated) using 20 µg/mL of Rabbit Anti-Human/Mouse STAT5a/b Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2168). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

• Reactivity: Hu, Mu
• Applications: WB, Simple Western, ChIP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: LYOPH

STAT5a/b Antibody [Unconjugated] - Pan Specific Summary

• Immunogen: Human STAT5a and STAT5b synthetic peptide
  aa 219-390
• Specificity: Detects endogenous human and mouse STAT5A and STAT5B in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: STAT5A
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP) 5 ug/5 x 10^6 cells
  • Simple Western 20 ug/mL
  • Western Blot 1 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for STAT5a/b Antibody [Unconjugated] - Pan Specific

• MGF
• STAT5
• STAT5A signal transducer and activator of transcription 5A
• STAT5a/b

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for STAT5a/b Antibody (AF2168)(14)

Publications using AF2168

• GM Wiedemann, EK Santosa, S Grassmann, S Sheppard, JB Le Luduec, NM Adams, C Dang, KC Hsu, JC Sun, CM Lau Deconvoluting global cytokine signaling networks in natural killer cells Nature Immunology, 2021;0(0):. 2021-01-01 [PMID: 33859404] (ChIP, Mouse)
• AM Schoepf, S Salcher, P Obexer, R Gust Tackling resistance in chronic myeloid leukemia: Novel cell death modulators with improved efficacy European Journal of Medicinal Chemistry, 2021;216(0):113285. 2021-01-01 [PMID: 33662676] (Western Blot, Human)
• GM Wiedemann, S Grassmann, CM Lau, M Rapp, AV Villarino, C Friedrich, G Gasteiger, JJ O'Shea, JC Sun Divergent Role for STAT5 in the Adaptive Responses of Natural Killer Cells Cell Reports, 2020;33(11):108498. 2020-01-01 [PMID: 33326784] (ChIP, Mouse)
• B Heizmann, S Le Gras, C Simand, P Marchal, S Chan, P Kastner Ikaros antagonizes DNA binding by STAT5 in pre-B cells PLoS ONE, 2020;15(11):e0242211. 2020-01-01 [PMID: 33180866] (ChIP, Mouse)
• W Elbezanti, A Lin, A Schirling, A Jackson, M Marshall, RV Duyne, F Maldarelli, L Sardo, Z Klase Benzodiazepines Drive Alteration of Chromatin at the Integrated HIV-1 LTR Viruses, 2020;12(2):. 2020-01-01 [PMID: 32050449] (Neutralization, Human)
• SH Yip, N Romanò, P Gustafson, DJ Hodson, EJ Williams, IC Kokay, AO Martin, P Mollard, DR Grattan, SJ Bunn Elevated Prolactin during Pregnancy Drives a Phenotypic Switch in Mouse Hypothalamic Dopaminergic Neurons Cell Rep, 2019;26(7):1787-1799.e5. 2019-01-01 [PMID: 30759390] (Western Blot, Mouse)
• SS Ganaie, W Zou, P Xu, X Deng, S Kleiboeker, J Qiu Phosphorylated STAT5 directly facilitates parvovirus B19 DNA replication in human erythroid progenitors through interaction with the MCM complex PLoS Pathog., 2017;13(5):e1006370. 2017-01-01 [PMID: 28459842] (Western Blot, Human)
• L Mokada-Gop, A Boeser, CH Lehmann, F Drepper, D Dudziak, B Warscheid, D Voehringer Identification of Novel STAT6-Regulated Proteins in Mouse B Cells by Comparative Transcriptome and Proteome Analysis J. Immunol, 2017;0(0):. 2017-01-01 [PMID: 28348271] (Western Blot, Mouse)
• Wagatsuma K, Tani-ichi S, Liang B, Shitara S, Ishihara K, Abe M, Miyachi H, Kitano S, Hara T, Nanno M, Ishikawa H, Sakimura K, Nakao M, Kimura H, Ikuta K STAT5 Orchestrates Local Epigenetic Changes for Chromatin Accessibility and Rearrangements by Direct Binding to the TCRgamma Locus. J Immunol, 2015;195(4):1804-14. 2015-01-01 [PMID: 26195811] (ChIP, Mouse)
• Richard A, Tan C, Hawley E, Gomez-Rodriguez J, Goswami R, Yang X, Cruz A, Penumetcha P, Hayes E, Pelletier M, Gabay O, Walsh M, Ferdinand J, Keane-Myers A, Choi Y, O'Shea J, Al-Shamkhani A, Kaplan M, Gery I, Siegel R, Meylan F The TNF-family ligand TL1A and its receptor DR3 promote T cell-mediated allergic immunopathology by enhancing differentiation and pathogenicity of IL-9-producing T cells. J Immunol, 2015;194(8):3567-82. 2015-01-01 [PMID: 25786692] (ChIP, Mouse)
• Zhang W, Gao C, Konopleva M, Chen Y, Jacamo R, Borthakur G, Cortes J, Ravandi F, Ramachandran A, Andreeff M Reversal of acquired drug resistance in FLT3-mutated acute myeloid leukemia cells via distinct drug combination strategies. Clin Cancer Res, 2014;20(9):2363-74. 2014-01-01 [PMID: 24619500] (Western Blot, Mouse)
• Liao W, Spolski R, Li P, Du N, West E, Ren M, Mitra S, Leonard W Opposing actions of IL-2 and IL-21 on Th9 differentiation correlate with their differential regulation of BCL6 expression. Proc Natl Acad Sci U S A, 2014;111(9):3508-13. 2014-01-01 [PMID: 24550509] (ChIP, Mouse)
• Zhang , Qian, Wang , Hong Yi, Wei , Fang, Liu , Xiaobin, Paterson , Jennifer, Roy , Darshan, Mihova , Daniela, Woetmann , Anders, Ptasznik , Andrzej, Odum , Niels, Schuster , Stephen, Marafioti , Teresa, Riley , James L, Wasik , Mariusz Cutaneous T cell lymphoma expresses immunosuppressive CD80 (B7-1) cell surface protein in a STAT5-dependent manner. J Immunol, 2014;192(6):2913-9. 2014-01-01 [PMID: 24523507] (ChIP, Human)
• Zhu J, Guo L, Min B, Watson CJ, Hu-Li J, Young HA, Tsichlis PN, Paul WE Growth factor independent-1 induced by IL-4 regulates Th2 cell proliferation. Immunity, 2002;16(5):733-44. 2002-01-01 [PMID: 12049724] (Western Blot, Mouse)

Bioinformatics

• Gene Symbol: STAT5A