Phospho-STAT1 (Tyr701) Monoclonal Antibody (KIKSI0803), eFluor™ 660, eBioscience™, Invitrogen™

Description

Description: This KIKSI0803 monoclonal antibody recognizes signal transducer and activator of transcription 1 (STAT1) when phosphorylated on tyrosine 701. STAT proteins are activated by ligand binding to receptors, such as cytokine receptors, that associate with Janus kinase (JAK) family members. Following their phosphorylation by JAKs, STAT proteins translocate to the nucleus where they bind to DNA and regulate transcription of specific genes in a cell type- and cytokine-specific manner. Phosphorylation of STAT1 on tyrosine 701 by JAK1 and JAK2 is essential for STAT1 dimer formation, nuclear translocation, and DNA binding activity. In response to IFN gamma stimulation, STAT1 homodimerizes or forms heterodimers with STAT3 that can bind to GAS (IFN gamma-activated sequence) promoter elements. In response to either IFN alpha or IFN beta stimulation, STAT1 forms a heterodimer with STAT2 that can bind ISRE (IFN-stimulated response element) promoter elements. Specificity of this KIKSI0803 clone was determined by ELISA, flow cytometry, and western blotting. Applications Reported: This KIKSI0803 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This KIKSI0803 antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of stimulated normal human peripheral blood cells. This can be used at 5 μL (0.06 μg) per test.

A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. Staining Protocol: We recommend using Protocol C: Two-step protocol: Fixation/Methanol. Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins and Protocol B: One-step protocol: intracellular (nuclear) proteins cannot be used. All Protocols can be found in the Flow Cytometry Protocols: "Staining Intracellular Antigens for Flow Cytometry Protocol" located in the Best Protocols Section under the Resources tab online. eFluor™ 660 is a replacement for Alexa Fluor™ 647. eFluor™ 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser. Filtration: 0.2 μm post-manufacturing filtered. STAT1 (signal transducers and activators of transcription 1) is a member of the STAT family of transcription factors. STAT1 can be activated by interferon-alpha, interferon-gamma, EGF, PDGF and IL6. STAT1 is known to regulate several genes which are involved in cell growth, apoptosis, immune responses, and lipid metabolism. Further, STAT1 plays an important role in mediating cell viability in response to different cell stimuli and pathogen exposure. The STAT1 gene is located on chromosome 2. STAT1 is activated to regulate gene expression in response to extracellular signaling polypeptides including cytokines, interferons, and growth factors. After phosphorylation by JAK tyrosine kinases, STAT1 enters the nucleus to regulate transcription of many different genes. Among the seven STATs types, STAT1, STAT3, STAT5a, and STAT5b have a wide activation profile. STAT1 is activated by many different ligands including IFN family (IFN-Alpha, IFN-Beta, IFN-gamma and IL-10), gp130 family (IL-6, IL-11, LIF, CNTF, and G-CSF), and receptor tyrosine kinases (EGF, PDGF, and CSF-1). Two alternatively spliced transcript variants encoding distinct isoforms of STAT1 have been described.