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

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

Catalog #: AF4190
3 Citations Datasheet / COA / SDS

Discontinued Product

AF4190 has been discontinued.

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

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Product Details

Citations (3)

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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

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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.

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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

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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

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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.

3 Citations: Showing 1 - 3
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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)
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