APC Mouse anti-Stat3
Clone M59-50 (RUO)
- Brand BD Phosflow™
- Alternative Name STAT3; Acute-phase response factor; APRF; HIES
- Vol. Per Test 20 µl
- Isotype Mouse BALB/c IgG1, λ
- Reactivity Human (QC Testing) Mouse, Rat (Predicted)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Human Stat3 Recombinant Protein
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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. 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. Upon activation, Stat3 dimerizes, translocates to the nucleus and binds DNA response elements, thereby regulating gene expression. It has been reported that Stat3 binds to DNA as a homodimer, but it is also capable of binding as a heterodimer with Stat1. 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-γ induced genes.
The M59-50 monoclonal antibody recognizes Stat3 (isoform 1) regardless of phosphorylation status. The specificity of this antibody conjugate for flow cytometric analysis was validated by confirming that RNA-mediated interference (RNAi) of the specific protein reduced the staining of the cells (see figure). Furthermore, the capacity of the RNAi to down-regulate the expression of the relevant protein was confirmed by western blot analysis.
Allophycocyanin (APC), is an accessory photosynthetic pigment found in blue-green algae. Its molecular weight is approximately 105 kDa. APC has six phycocyanobilin chromophores per molecule, which makes it a very bright fluorochrome that is highly suitable for flow cytometry applications. 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 APC under optimum conditions, and unconjugated antibody and free APC were 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
This antibody conjugate is suitable for intracellular staining of human cell lines and peripheral blood mononuclear cells using BD Cytofix™ Fixation Buffer. Either BD Phosflow™ Perm Buffer II or Perm Buffer III may be used. We have observed that BD Phosflow™ Perm/Wash Buffer I gives unsatisfactory results with this antibody.