Human Phospho-STAT5a/b (Y694/Y699) Antibody
R&D Systems, part of Bio-Techne | Catalog # AF4190
Discontinued Product
AF4190 has been discontinued.
View all STAT5a/b products.
Key Product Details
Species Reactivity
Validated:
Human
Cited:
Human, Mouse
Applications
Validated:
Western Blot, Intracellular Staining by Flow Cytometry, Immunocytochemistry, Simple Western, CyTOF-ready
Cited:
Western Blot, Chromatin Immunoprecipitation (ChIP), ELISA Development (Capture)
Label
Unconjugated
Antibody Source
Polyclonal Rabbit IgG
Product Specifications
Immunogen
Synthetic phosphopeptide containing human STAT5b Y699 site (aa sequence of this peptide is identical to a correspondent region of human STAT5a containing Y694).
Specificity
Detects human STAT5a/b when phosphorylated at Y694/Y699 in Western blots.
Clonality
Polyclonal
Host
Rabbit
Isotype
IgG
Scientific Data Images for Human Phospho-STAT5a/b (Y694/Y699) Antibody
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.
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.
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.Applications for Human Phospho-STAT5a/b (Y694/Y699) Antibody
Application
Recommended Usage
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
Sample: Immersion fixed IFN-alpha treated Daudi human Burkitt's lymphoma cell line
Sample: Immersion fixed IFN-alpha treated Daudi human Burkitt's lymphoma cell line
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
Sample: Daudi human Burkitt's lymphoma cell line fixed with paraformaldehyde and permeabilized with ice-cold methanol
Sample: Daudi human Burkitt's lymphoma cell line fixed with paraformaldehyde and permeabilized with ice-cold methanol
Simple Western
10 µg/mL
Sample: Daudi human Burkitt's lymphoma cell line treated with Recombinant Human IFN‑ alphaA (Catalog # 11100-1)
Sample: Daudi human Burkitt's lymphoma cell line treated with Recombinant Human IFN‑ alphaA (Catalog # 11100-1)
Western Blot
1 µg/mL
Sample: Daudi human Burkitt's lymphoma cell line and U937 human histiocytic lymphoma cell line treated with Human Interferon Alpha A/D [BglII] (Catalog # 11200-1)
Sample: Daudi human Burkitt's lymphoma cell line and U937 human histiocytic lymphoma cell line treated with Human Interferon Alpha A/D [BglII] (Catalog # 11200-1)
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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.
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
Additional STAT5a/b Products
Product Documents for Human Phospho-STAT5a/b (Y694/Y699) Antibody
Product Specific Notices for Human Phospho-STAT5a/b (Y694/Y699) Antibody
For research use only
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