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Alexa Fluor® 488 Mouse Anti-Stat4 (pY693)
Alexa Fluor® 488 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-α 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 usingBD Phosflow™ Perm Buffer III, (Cat. No. 558050) on ice for at least 30 minutes, and then stained with Alexa Fluor® 488 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-α 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 usingBD Phosflow™ Perm Buffer III, (Cat. No. 558050) on ice for at least 30 minutes, and then stained with Alexa Fluor® 488 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 (QC Testing), Mouse (Tested in Development)
Mouse IgG2b, κ
Phosphorylated Human Stat4 (pY693)
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
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 to Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was 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.
  6. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  7. 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.
  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.
558136 Rev. 3
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.

Clone 38/p-Stat4 has been confirmed to recognize mouse phospho-Stat4 with western blot application using Purified Mouse Anti-Stat4 (pY693), Cat. Nos. 612739 & 612738.

558136 Rev. 3
Format Details
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Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
558136 Rev.3
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).
558136 Rev. 3

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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.