Human Phospho-STAT5a/b (Y694/Y699) Antibody: R&D Systems

Human Phospho-STAT5a/b (Y694/Y699) Antibody

AF4190 has been discontinued. View all STAT5a/b products.
(3 citations)   
  • Species Reactivity
  • Specificity
    Detects human STAT5a/b when phosphorylated at Y694/Y699 in Western blots.
  • Source
    Polyclonal Rabbit IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Synthetic phosphopeptide containing human STAT5b Y699 site (aa sequence of this peptide is identical to a correspondent region of human STAT5a containing Y694).
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
  • Western Blot
    1 µg/mL
    See below
  • Simple Western
    10 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    2.5 µg/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Intracellular Staining by Flow Cytometry
Detection of STAT5a/b in Daudi Human Cell Line by Flow Cytometry. Daudi human Burkitt's lymphoma cell line was unstimulated (light orange filled histogram) or treated with 500 U/mL Recombinant Human IFN-alpha (Catalog # 11100-1) for 20 minutes, then stained with Rabbit Anti-Human Phospho-STAT5a/b (Y694/Y699) Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF4190, dark orange filled histogram) or isotype 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 ice-cold methanol.
STAT5a/b in Daudi Human Burkitt's Lymphoma Cells. STAT5a/b phosphorylated at Y694/Y699 was detected in immersion fixed IFN-alpha treated Daudi human Burkitt's lymphoma cell line using 10 µg/mL Rabbit Anti-Human Phospho-STAT5a/b (Y694/Y699) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4190) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red, upper panel; Catalog # NL004) and counterstained with DAPI (blue, lower panel). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human Phospho-STAT5a/b (Y694/Y699) by Western Blot. Western blot shows lysates of Daudi human Burkitt's lymphoma cell line and U937 human histiocytic lymphoma cell line untreated (-) or treated (+) with 500 U/mL Recombinant Human
IFN‑ alpha  2a (Catalog # 11100-1) for 15 minutes. Gels were loaded with 20 μg of cytoplasmic (Cyto) and 10 μg of nuclear extracts (Nuc). PVDF membrane was probed with 1 µg/mL Rabbit Anti-Human Phospho-STAT5a/b (Y694/Y699) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4190) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band for Phospho-STAT5a/b (Y694/Y699) was detected at approximately 105 kDa (as indicated). For additional reference, duplicate lysates were probed with 1.0  µg/mL Human/Mouse STAT5a/b Pan Specific Antigen Affinity-purified Polyclonal Antibody (lower panel, Catalog # AF2168). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Phospho-STAT5a/b (Y694/Y699) by Simple WesternTM.

Simple Western lane view shows lysates of Daudi human Burkitt's lymphoma cell line untreated (-) or treated (+) with 500 U/mL Recombinant Human IFN‑ alpha A (Catalog # 11100-1) for 15 minutes, loaded at 0.2 mg/mL. A specific band was detected for STAT5a/b at approximately 68 kDa (as indicated) using 10 µg/mL of Rabbit Anti-Human Phospho-STAT5a/b (Y694/Y699) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4190). 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.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & 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.
Background: STAT5a/b

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 (IL‑22, IL-6, IFN-alpha ). 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.

  • Long Name:
    Signal Transducer and Activator of Transcription 5b
  • Alternate Names:
    MGF; STAT5; STAT5A signal transducer and activator of transcription 5A ; STAT5a/b
Related Research Areas

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.

3 Citations: Showing 1 - 3
Filter your results:

Sample Type
  1. MiR-21 is under control of STAT5 but is dispensable for mammary development and lactation.
    Authors: Feuermann, Yonatan, Kang, Keunsoo, Shamay, Avi, Robinson, Gertraud, Hennighausen, Lothar
    PLoS ONE, 2014;9(1):e85123.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: ChIP
  2. Erythropoietin production in neuroepithelial and neural crest cells during primitive erythropoiesis.
    Authors: Suzuki N, Hirano I, Pan X, Minegishi N, Yamamoto M
    Nat Commun, 2013;4(0):2902.
    Species: Mouse
    Sample Type: Whole Cells
    Application: ELISA Capture
  3. Colocalization of the VEGF-R2 and the common IL-3/GM-CSF receptor beta chain to lipid rafts leads to enhanced p38 activation.
    Authors: Saulle E, Riccioni R, Coppola S, Parolini I, Diverio D, Riti V, Mariani G, Laufer S, Sargiacomo M, Testa U
    Br. J. Haematol., 2009;145(3):399-411.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB

Have you used Human Phospho-STAT5a/b (Y694/Y699) Antibody?

Submit a review and receive a $25US/€18/£15/$25CAN amazon gift card if you include an image - $10US/€7/£6/$10CAN Amazon card for reviews without an image. Limited to verified customers in USA, Canada and Europe.

Order Details

Contact Information


Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

R&D Systems Guarantee

Panels For

Learn More
Monoclonal Antibodies


Customer Information