Alexa Fluor® 647 Mouse Anti-Stat3 (pS727)
Clone 49/p-Stat3 (RUO)
- Brand BD Phosflow™
- Vol. Per Test 20 µl
- Isotype Mouse IgG1
- Reactivity Human (QC Testing) Mouse, Rat (Tested in Development)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Phosphorylated Human Stat3
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The 49/p-Stat3 monoclonal antibody recognizes the S727-phosphorylated form of Signal transducer and activator of transcription 3 (Stat3/STAT3 isoform 1). The fluorochrome-conjugated formats have been evaluated using a human model system. However, the unconjugated form of this antibody is also effective for western blot analysis of human, mouse, and rat tissue. The Stat proteins function both as cytoplasmic signal transducers and as activators of transcription. Seven mammalian Stat proteins have been identified: Stat1-4, Stat5a, 5b, and Stat6. Stat3 is a 92-kDa protein that is activated as a DNA binding protein through cytokines, such as IL-6, and growth factors, such as EGF. Stat3 is phophorylated at serine 727 (S727) via the MAPK pathway. The S727 residue is located at a conserved Pro-X-Ser-Pro sequence, which is recognized by the protein kinase ERK. Activation through the S727 residue is thought to lead to initiation of transcription. Upon activation, Stat3 dimerizes, translocates to the nucleus, and binds DNA response elements thereby regulating gene expression. It appears that Stat3 binds to DNA as a homodimer, but it is also capable of binding as a heterodimer with Stat1. In addition to serine phosphorylation, Stat3 is also phosphorylated at tyrosine 705 by JAK1 in response to cytokine stimulation. Stat3 is widely expressed and can bind to the sis-inducible element (SIE) site from the c-fos promoter. This site is similar to the GAS element that is present in IFN--induced genes. Thus, phosphorylation of S727 in Stat3 occurs in response to growth factors and cytokines, and is essential for normal transcriptional activity.
Alexa Fluor® 647 conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of APC. However, APC tends to be brighter while Alexa Fluor® 647 is more optimal for intracellular applications. This fluorochrome exhibits uncommon photostability, making it an ideal choice for use in fluorescence microscopy. Due to nearly identical excitation and emission properties but different spillover characteristics, APC and Alexa Fluor® 647 cannot be used simultaneously.
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Preparation and Storage
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.
- 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).
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.