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 PerCP-CY™5.5 anti-Stat6 (pY641) antibody (Cat. No. 561195). 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 PerCP-CY™5.5 anti-Stat6 (pY641) antibody (Cat. No. 561195). 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.
製品詳細
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_10563081
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 with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.
Product Notices
- 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).
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
- PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
関連製品
Stain Buffer (BSA) RUO
サイズ
500 mL
カタログ番号
554657
Lyse/Fix Buffer 5X RUO
サイズ
250 mL
カタログ番号
558049
Perm Buffer III RUO
サイズ
125 mL
カタログ番号
558050
Recombinant Human IL-4 RUO
サイズ
5 µg
カタログ番号
554605
561195 Rev. 2
抗体の詳細
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.
561195 Rev. 2
フォーマットの詳細
PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
561195 Rev.2
引用＆参考文献
View product citations for antibody "561195" on CiteAb
Development References (6)
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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
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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
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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
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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
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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
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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
561195 Rev. 2
Please refer to Support Documents for Quality Certificates
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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.