Description
Stat (Signal transducer and activators of transcription) proteins are critical mediators of the biologic activity of cytokines, including interleukins, interferons, erythropoietin, and growth factors. Ligand-receptor interaction leads to activation of constitutively associated JAK family kinases and subsequent recruitment/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, which is the primary transcription activator induced by the binding of the interferon 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 is translocated to the nucleus. This results in the formation of an active complex that includes the DNA-binding p48 subunit. This complex is responsible for modulating the transcription of the interferon-stimulated genes (ISGs). Thus, phosphorylation of Y701 in Stat1 occurs in response to growth factors and cytokines, and is essential for normal transcriptional activity of the ISGF3 complex. The 1/Stat1 monoclonal antibody recognizes the N-terminus of human Stat1 (both isoforms), regardless of phosphorylation status.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
2. An isotype control should be used at the same concentration as the antibody of interest.
3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
6. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
8. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
9. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
