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Alexa Fluor® 647 Mouse Anti-Stat6 (pY641)
Alexa Fluor® 647 Mouse Anti-Stat6 (pY641)

Flow cytometric analysis of phospho-Stat6 (pY641). Human endothelial cells were starved overnight in DMEM containing 0.1% FBS. The following day, cells were either left unstimulated (open diagram) or stimulated (shaded) with human IL-4 recombinant protein (Cat. No. 554605) at 10 ng/ml for 60 minutes at 37°C. Cells were fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655) for 10 minutes at 37°C and then permeabilized in BD Phosflow™ Perm Buffer III (Cat. No. 558050) for 30 minutes on ice or overnight at -20°C. Cells were then washed twice in BD Pharmingen™ Stain Buffer (Cat. No. 554656), and stained with Alexa Fluor® 647-conjugated phospho-Stat6 (pY641) antibody for 1 hour at RT, (Cat. No. 612601). The cells were analyzed on a BD FACSCalibur™ flow cytometer.

Flow cytometric analysis of phospho-Stat6 (pY641). Human endothelial cells were starved overnight in DMEM containing 0.1% FBS. The following day, cells were either left unstimulated (open diagram) or stimulated (shaded) with human IL-4 recombinant protein (Cat. No. 554605) at 10 ng/ml for 60 minutes at 37°C. Cells were fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655) for 10 minutes at 37°C and then permeabilized in BD Phosflow™ Perm Buffer III (Cat. No. 558050) for 30 minutes on ice or overnight at -20°C. Cells were then washed twice in BD Pharmingen™ Stain Buffer (Cat. No. 554656), and stained with Alexa Fluor® 647-conjugated phospho-Stat6 (pY641) antibody for 1 hour at RT, (Cat. No. 612601). The cells were analyzed on a BD FACSCalibur™ flow cytometer.

Product Details
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BD Phosflow™
Signal transducer and activator of transcription 6; IL-4 Stat
Human (QC Testing)
Mouse IgG2a
Phosphorylated Human Stat6 (pY641)
Intracellular staining (flow cytometry) (Routinely Tested)
100 kDa
20 µl
Aqueous buffered solution containing BSA 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® 647 under optimum conditions, and unreacted Alexa Fluor® 647 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. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  4. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  5. 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.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
612601 Rev. 4
Antibody Details
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18/P-Stat6

Interleukin-4 (IL-4), a major immunoregulatory cytokine, is secreted by activated T lymphocytes, basophils, and mast cells and plays an important role in modulating T helper cell lineage development. It induces specific gene expression via the tyrosine phosphorylation of Stat6 at tyrosine 641 (Y641). Stat6, a member of the signal transducers and activators of transcription protein family, mediates signals for IL-4 and, possibly, IL-13. While Stat6 is widely expressed in human tissues, it exhibits elevated expression in peripheral blood lymphocytes, colon, intestine, ovary, prostate, thymus, spleen, kidney, liver, lung, and placenta. Following cytokine receptor ligation, Jak kinases are activated and phosphorylate the cytoplasmic tails of the oligomerized receptors. The SH3:SH2 domain of Stat6 associates with tyrosine-phosphorylated IL-4 receptor and the proximal Jak kinase phosphorylates Stat6 at Y641 on the C-terminal side of the SH2 domain. Stat6 is then released from the receptor, dimerizes, and is thought to contact the basal transcription machinery by binding to p300/CBP. Thus, Stat6 mediates the IL-4 signal and is essential for the proper development of adaptive immunity.

612601 Rev. 4
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
612601 Rev.4
Citations & References
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Development References (6)

  1. Bromberg J, Darnell JE. The role of STATs in transcriptional control and their impact on cellular function. Oncogene. 2000; 19(21):2468-2473. (Biology). View Reference
  2. Dent AL, Hu-Li J, Paul WE, Staudt LM. T helper type 2 inflammatory disease in the absence of interleukin 4 and transcription factor STAT6. Proc Natl Acad Sci U S A. 1998; 95(23):13823-13828. (Biology). View Reference
  3. Heim MH. The Jak-STAT pathway: specific signal transduction from the cell membrane to the nucleus. Eur J Clin Invest. 1996; 26(1):1-12. (Biology). View Reference
  4. Hou J, Schindler U, Henzel WJ, Ho TC, Brasseur M, McKnight SL. An interleukin-4-induced transcription factor: IL-4 Stat. Science. 1994; 265(5179):1701-1706. (Biology). View Reference
  5. Mikita T, Campbell D, Wu P, Williamson K, Schindler U. Requirements for interleukin-4-induced gene expression and functional characterization of Stat6. Mol Cell Biol. 1996; 16(10):5811-5820. (Biology). View Reference
  6. Quelle FW, Shimoda K, Thierfelder W, et al.. Cloning of murine Stat6 and human Stat6, Stat proteins that are tyrosine phosphorylated in responses to IL-4 and IL-3 but are not required for mitogenesis. Mol Cell Biol. 1995; 15(6):3336-3343. (Biology). View Reference
View All (6) View Less
612601 Rev. 4

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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.