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PE Mouse Anti-Stat4 (pY693)
PE Mouse Anti-Stat4 (pY693)

Analysis of Stat4 (pY693) in activated human peripheral blood lymphocytes.  Human whole blood was either stimulated with 40,000 U/ml of IFN-a for 15 minutes at 37ºC (open histogram) or unstimulated (shaded histogram).  The cells were lysed and fixed with 1X BD PhosFlow™ Lyse/Fix Buffer (Cat. No. 558049) for 10-15 minutes at 37ºC, then permeabilized (BD PhosFlow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with PE anti-Stat4 (pY693).  For data analysis, lymphocytes were selected by scatter profile.  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Analysis of Stat4 (pY693) in activated human peripheral blood lymphocytes.  Human whole blood was either stimulated with 40,000 U/ml of IFN-a for 15 minutes at 37ºC (open histogram) or unstimulated (shaded histogram).  The cells were lysed and fixed with 1X BD PhosFlow™ Lyse/Fix Buffer (Cat. No. 558049) for 10-15 minutes at 37ºC, then permeabilized (BD PhosFlow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with PE anti-Stat4 (pY693).  For data analysis, lymphocytes were selected by scatter profile.  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Product Details
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BD Phosflow™
Signal transducer and activator of transcription 4; SLEB11
Human, Mouse (QC Testing)
Mouse IgG2b, κ
Phosphorylated Human Stat4 (pY693)
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_10895804
Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.
RUO


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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Product Notices

  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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
562073 Rev. 1
Antibody Details
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38/p-Stat4

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.  Stat4 has been shown to play an important role in development of T helper cells, specifically the Th1 subset. Stat4 is activated by IL-12 and by type 1 interferons. Knockout mice supported the role that Stat4 plays in IL-12 signaling because lymphocytes from Stat 4-/- mice could neither differentiate into Th1 cells nor produce IFN-γ in response to treatment with IL-12. IFN-γ plays an important role in host defense. A key component in the activation of Stat4 is the phosphorylation on tyrosine and serine residues in response to IL-12 stimulation. IL-12 stimulation leads to the phosphorylation of Stat4 on tyrosine 693 and serine 721. Transcriptional activity of Stat4 has been shown to be significantly reduced when residues Y693 and S721 are mutated.

562073 Rev. 1
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
562073 Rev.1
Citations & References
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Development References (2)

  1. Kisseleva T, Bhattacharya S, Braunstein J, Schindler CW. Signaling through the JAK/STAT pathway, recent advances and future challenges. Gene. 2002; 285:1-24. (Biology).
  2. Visconti R, Gadina M, Chiariello M, et al. Importance of the MKK6/p38 pathway for interleukin-12−induced STAT4 serine phosphorylation and transcriptional activity. Blood. 2000; 96:1844-1852. (Biology).
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Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.