STAT5b Antibody (389215) [Unconjugated]

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, Daudi human Burkitt's lymphoma cell line, and K562 human chronic myelogenous leukemia cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human STAT5b Monoclonal Antibody (Catalog # MAB1584) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). 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.

Jurkat human acute T cell leukemia cell line was stained with Mouse Anti-Human STAT5b Monoclonal Antibody (Catalog # MAB1584, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0101B).

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for STAT5b at approximately 91 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human STAT5b Monoclonal Antibody (Catalog # MAB1584) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and STAT5b knockout HeLa cell line (KO). PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human STAT5b Monoclonal Antibody (Catalog # MAB1584) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for STAT5b at approximately 90 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunoprecipitation was performed on cell lysate of HAP1 human near-haploid cell line using 2.0 μg of Mouse Anti-Human Stat5b Monoclonal Antibody (Catalog # MAB1584) pre-coupled to protein G or protein A beads. Immunoprecipitated Stat5b was detected with a Rabbit 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).

• Reactivity: Hu
• Applications: WB, Simple Western, IP, CyTOF-ready, Flow, KO
• Clone: 389215
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

STAT5b Antibody (389215) [Unconjugated] Summary

• Immunogen: Human STAT5b synthetic peptide
  MDSQWIPHAQS
  Accession # NP_036580
• Specificity: Detects human STAT5b in Western blots.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: STAT5B
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Immunoprecipitation 2 ug/1 mg cell lysate
  • Intracellular Staining by Flow Cytometry 2.5 ug/10^6 cells
  • Knockout Validated
  • Simple Western 20 ug/mL
  • Western Blot 1 ug/mL

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 0.5 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 STAT5b Antibody (389215) [Unconjugated]

• signal transducer and activator of transcription 5B
• STAT5
• STAT5b
• transcription factor STAT5B

Background

Signal Transduction and Activator of Transcription 5B (STAT5b) is a member of the Jak/STAT signal transduction pathway and is activated by a variety of cytokines (IL-22, IL-6, IFN alpha ). Upon cytokine activation, STAT5b is phosphorylated and translocates to the nucleus where it binds to the DNA consensus site TTCN₃GAA and activates transcription of specific genes. STAT5b shares 93% amino acid identity with STAT5a. STAT5b 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 STAT5b Antibody (MAB1584)(1)

Publication using MAB1584

• Christopher J Cardinale, Michael E March, Xiang Lin, Yichuan Liu, Lynn A Spruce, Jonathan P Bradfield et al. Regulation of Janus Kinase 2 by an Inflammatory Bowel Disease Causal Non-coding Single Nucleotide Polymorphism Journal of Crohn's and Colitis 2020-06-19 [PMID: 32271392]

Review for STAT5b Antibody (MAB1584) (1)

Average Rating: 3

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer Simple Western Human 11/28/2016

Summary

• Application: Simple Western
• Sample Tested: k562 cells
• Species: Human

Comments

• Comments: Charge Assay – Stat5b, Novus Biologicals, MAB1584
  
  Theoretical pI = 5.71; no peak close to that value in all three replicates
  
  Cells used = K562 (human) lysed in Protein Simple bicine/CHAPS buffer (040-764) loaded at 100 ng/µL; Gradient = 5 – 8 nested, ladder# 3
  
  Primary antibody at 1:50
  
  Secondary antibody = Protein Simple biotinylated anti-mouse (041-127) with Protein Simple streptavidin-HRP (041-106) both at 1:100
  
  Size Assay – Stat5b, Novus Biologicals, MAB1584
  
  Predicted m.w. = 89.9 kDa; well defined peak at 93 kDa, good agreement between replicates
  
  Cells used = K562 (human) lysed in Protein Simple bicine/CHAPS buffer (040-764) loaded at 500 ng/µL
  
  Primary antibody at 1:50

Bioinformatics

• Gene Symbol: STAT5B
• Uniprot:
  • NP_036580()