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![RB780 Mouse Anti-Stat1 (pY701)](/content/dam/bdb/products/global/reagents/flow-cytometry-reagents/research-reagents/single-color-antibodies-ruo/569xxx/5691xx/569144_base/569144_569145-1a.png)
![RB780 Mouse Anti-Stat1 (pY701)](/content/dam/bdb/products/global/reagents/flow-cytometry-reagents/research-reagents/single-color-antibodies-ruo/569xxx/5691xx/569144_base/569144_569145-1a.png)
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![RB780 Mouse Anti-Stat1 (pY701)](/content/dam/bdb/products/global/reagents/flow-cytometry-reagents/research-reagents/single-color-antibodies-ruo/569xxx/5691xx/569144_base/569144_569145-1a.png)
Flow cytometric analysis of Stat1 (pY701) expression in Unstimulated and Stimulated Human CD4+ T lymphocytes. Human peripheral blood leucocytes were either not stimulated (Unstimulated, dashed line histogram) or stimulated (Stimulated, solid line histogram) for 15 minutes with Phorbol 12-Myristate 13-Acetate (Sigma P-8139) as well as Recombinant Human IFN-α, IL-2, IL-4, and IL-6 proteins. The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656], fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), and then permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). After washing and resuspending in Stain Buffer, the cells were stained with APC Mouse Anti-Human CD4 antibody (Cat. No. 561841) and BD Phosflow™ RB780 Mouse Anti-Stat1 (pY701) antibody. The fluorescence histograms showing Stat1 (pY701) expression were derived from gated CD4+ T cells with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD™ LSR II Flow Cytometry System and FlowJo™ software.
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![RB780 Mouse Anti-Stat1 (pY701)](/content/dam/bdb/products/global/reagents/flow-cytometry-reagents/research-reagents/single-color-antibodies-ruo/569xxx/5691xx/569144_base/569144_569145-1a.png)
BD Phosflow™ RB780 Mouse Anti-Stat1 (pY701)
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Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
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Stat (Signal transducer and activators of transcription) proteins mediate the biological activity of cytokines, including interleukins, interferons, erythropoietin, and growth factors. Ligand-receptor interaction activates constitutively-associated JAK family kinases as well as subsequent recruitment and activation of Stat proteins by tyrosine phosphorylation. Active Stat proteins then move to the nucleus to promote transcription of cytokine-inducible genes. Seven Stat proteins have been cloned, each of which is differentially expressed and/or activated in a cytokine-specific and cell type-specific manner. Stat1 and Stat2 are components of the ISGF3 (Interferon-Stimulated Gene Factor 3) complex, the primary transcription activator induced by interferon binding to a specific cell-surface receptor. Stat1 has two alternatively spliced isoforms, 91-kDa Stat1α and 84-kDa Stat1β; Stat1α has 38 additional C-terminal amino acids. In response to the binding of IFNα, IFNγ, EGF, PDGF, or CSF-1 to their respective receptors, the Stat1 subunits become tyrosine-phosphorylated at Y701, and the complex translocates to the nucleus. This forms an active complex that includes the DNA-binding p48 subunit, and is responsible for modulating interferon-stimulated genes (ISGs) transciption.
The 4a monoclonal antibody recognizes the phosphorylated Y701 in Stat1α and Stat1β.
![altImg](/content/dam/bdb/products/vialimages/packaging-images/formatimages/RB780_Spectrum.png)
Development References (6)
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Bromberg J, Darnell JE. The role of STATs in transcriptional control and their impact on cellular function. Oncogene. 2000; 19(21):2468-2473. (Biology). View Reference
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Darnell JE Jr. STATs and gene regulation. Science. 1997; 277(5332):1630-1635. (Biology). View Reference
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Fu XY, Zhang JJ. Transcription factor p91 interacts with the epidermal growth factor receptor and mediates activation of the c-fos gene promoter. Cell. 1993; 74(6):1135-1145. (Biology). View Reference
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Perez OD, Mitchell D, Campos R, Gao GJ, Li L, Nolan GP. Multiparameter analysis of intracellular phosphoepitopes in immunophenotyped cell populations by flow cytometry. Curr Protoc Cytom. 2005; 6.20.1-6.20.22. (Clone-specific: Flow cytometry). View Reference
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Suni MA, Maino VC. Flow cytometric analysis of cell signaling proteins. Methods Mol Biol. 2011; 717:155-169. (Clone-specific). View Reference
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Tanaka S, Saito Y, Kunisawa J, et al. Development of mature and functional human myeloid subsets in hematopoietic stem cell-engrafted NOD/SCID/IL2rgammaKO mice. J Immunol. 2012; 188(12):6145-6155. (Clone-specific: Flow cytometry). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.