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

Name: Human Phospho-STAT5a/b (Y694/Y699) Antibody
Brand: R&D Systems
SKU: AF4190

Catalog #: AF4190
4 Citations Datasheet / COA / SDS

Discontinued Product

AF4190 has been discontinued.

View all STAT5a/b products.

Detection of Human Phospho-STAT5a/b (Y694/Y699) by Western Blot.

4 Images

All STAT5a/b Antibodies

Product Details

Citations (4)

FAQs

Reviews

Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

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

Species Reactivity

Human

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

Unconjugated

Applications

Recommended Concentration

Sample

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.

Scientific Data

Western Blot

View Larger

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-a 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.

Intracellular Staining by Flow Cytometry

Detection of STAT5a/b antibody in Daudi Human Cell Line antibody by Flow Cytometry.

View Larger

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-a (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.

Immunocytochemistry

View Larger

STAT5a/b in Daudi Human Burkitt's Lymphoma Cells. STAT5a/b phosphorylated at Y694/Y699 was detected in immersion fixed IFN-a 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.

Simple Western

View Larger

Detection of Human Phospho-STAT5a/b (Y694/Y699) by Simple Western^TM. Simple Western lane view shows lysates of Daudi human Burkitt's lymphoma cell line untreated (-) or treated (+) with 500 U/mL Recombinant Human IFN-aA (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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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

Product Datasheets

Product Datasheet

COA

SDS

Related Research Areas

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

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.

4 Citations: Showing 1 - 4
Filter your results:

Filter by:

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, 2014-01-30;9(1):e85123.
Species: Mouse
Sample Types: Cell Lysates
Applications: ChIP
Erythropoietin production in neuroepithelial and neural crest cells during primitive erythropoiesis.
Authors: Suzuki N, Hirano I, Pan X, Minegishi N, Yamamoto M
Nat Commun, 2013-01-01;4(0):2902.
Species: Mouse
Sample Types: Whole Cells
Applications: ELISA Development (Capture)
Location and the functionality of erythropoietin receptor(s) in A2780 cells
Authors: Peter Solár, Gabriela Hrčková, Lenka Varinská, Zuzana Solárová, Ján Kriška, Ivana Uhrínová et al.
Oncology Reports
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot
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., 2009-02-24;145(3):399-411.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot