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V450 Mouse Anti-Stat6 (pY641)
V450 Mouse Anti-Stat6 (pY641)

Flow cytometric analysis of phospho-Stat6 (pY641) in human peripheral blood lymphocytes. Whole blood was either left untreated (dashed line histogram) or treated (solid line histogram) with 100 ng/ml (final concentration) of BD Pharmingen™ Recombinant Human IL-4 (Cat. No. 554605) for 15 minutes at 37°C. The samples were lysed and fixed with 1X BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) for 10 minutes at 37°C, permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) on ice for 30 minutes and were then stained with BD Horizon™ V450 anti-Stat6 (pY641) (Cat No. 561203). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Flow cytometric analysis of phospho-Stat6 (pY641) in human peripheral blood lymphocytes. Whole blood was either left untreated (dashed line histogram) or treated (solid line histogram) with 100 ng/ml (final concentration) of BD Pharmingen™ Recombinant Human IL-4 (Cat. No. 554605) for 15 minutes at 37°C. The samples were lysed and fixed with 1X BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) for 10 minutes at 37°C, permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) on ice for 30 minutes and were then stained with BD Horizon™ V450 anti-Stat6 (pY641) (Cat No. 561203). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

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)
5 µl
AB_10565979
Aqueous buffered solution containing 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 BD Horizon™ V450 under optimum conditions, and unreacted BD Horizon™ V450 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. 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.
  3. BD Horizon V450 has a maximum absorption of 406 nm and maximum emission of 450 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
  4. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
561203 Rev. 2
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.

The antibody is conjugated to BD Horizon™ V450, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser Ex max of 406 nm and has an Em Max at 450 nm. Conjugates with BD Horizon™ V450 can be used in place of Pacific Blue™ conjugates.

561203 Rev. 2
Format Details
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V450
BD Horizon™ V450 Dye is part of the BD Horizon™ violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 405-nm and an emission maximum (Em Max) at 450-nm. BD Horizon™ V450, driven by BD innovation, is designed to be excited by the violet laser (405 nm) and detected using an optical filter centered near 450-nm (e.g., a 450/50-nm bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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V450
Violet 405 nm
405 nm
450 nm
561203 Rev.2
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
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561203 Rev. 2

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.