Revision 5
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IF-IC, FC-FP, ChIP

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

110

Source/Isotype:

Rabbit IgG

UniProt ID:

#P42226

Entrez-Gene Id:

6778

Product Information

Product Usage Information

For optimal ChIP results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:200 - 1:800
Flow Cytometry (Fixed/Permeabilized) 1:50 - 1:200
Chromatin IP 1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #46186.

Specificity / Sensitivity

Phospho-Stat6 (Tyr641) (D8S9Y) Rabbit mAb recognizes endogenous levels of Stat6 protein only when phosphorylated at Tyr641. Weak cross-reactivity with other tyrosine-phosphorylated proteins has been observed in some cell lines.

Species Reactivity:

Human, Mouse

Species predicted to react based on 100% sequence homology

Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phospho-peptide corresponding to residues surrounding Tyr641 of human Stat6 protein.

Background

Upon activation by Janus kinases, Stat6 translocates to the nucleus where it regulates cytokine-induced gene expression. Stat6 is activated via phosphorylation at Tyr641 and is required for responsiveness to IL-4 and IL-13 (1-4). In addition, Stat6 is activated by IFN-α in B cells, where it forms transcriptionally active complexes with Stat2 and p48 (5,6). Protein phosphatase 2A is also involved in regulation of IL-4-mediated Stat6 signaling (7).

  1. Nelms, K. et al. (1999) Ann. Rev. Immunol. 17, 701-738.
  2. Malabarba, M.G. et al. (1996) Biochem. J. 319, 865-872.
  3. Hou, J. et al. (1994) Science 265, 1701-1706.
  4. Quelle, F.W. et al. (1995) Mol. Cell. Biol. 15, 3336-3343.
  5. Takeda, K. et al. (1996) Nature 380, 627-630.
  6. Gupta, S. et al. (1999) J. Immunol. 163, 3834-3841.
  7. Woetmann, A. et al. (2003) J. Biol. Chem. 278, 2787-2791.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 5
#56554

Phospho-Stat6 (Tyr641) (D8S9Y) Rabbit mAb

Western Blotting Image 1: Phospho-Stat6 (Tyr641) (D8S9Y) Rabbit mAb Expand Image
Western blot analysis of extracts from ACHN cells, untreated (-) or treated with Human Interleukin-4 (hIL-4) #8919 (100 ng/ml, 10 min; +), and BaF3 cells, untreated (-) or treated with Mouse Interleukin-3 (mIL-3) #8923 (10 ng/ml, 5 min; +), using Phospho-Stat6 (Tyr641) (D8S9Y) Rabbit mAb (upper), Stat (D3H4) Rabbit mAb #5397 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunoprecipitation Image 1: Phospho-Stat6 (Tyr641) (D8S9Y) Rabbit mAb Expand Image
Immunoprecipitation of phospho-Stat6 (Tyr641) from ACHN cells treated with Human Interleukin-4 (hIL-4) #8919 (10 ng/ml, 10 min). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-Stat6 (Tyr641) (D8S9Y) Rabbit mAb. Western blot was performed using Phospho-Stat6 (Tyr641) (D8S9Y) Rabbit mAb.
Immunofluorescence Image 1: Phospho-Stat6 (Tyr641) (D8S9Y) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of ACHN cells (Stat6 high-expressing), serum starved (left) or treated with Human Interleukin-4 (hIL-4) #8919 (10 ng/mL, 10 min; center), and LNCaP cells (Stat6 low-expressing) treated with hIL-4 (right) using Phospho-Stat6 (Tyr641) (D8S9Y) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red).
Flow Cytometry Image 1: Phospho-Stat6 (Tyr641) (D8S9Y) Rabbit mAb Expand Image
Flow cytometric analysis of ACHN cells, untreated (blue) or treated with hIL-4 (10 ng/mL, 10 min; green), using Phospho-Stat6 (Tyr641) (D8S9Y) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 fragment (Alexa Fluor® 488 conjugate) #4412 was used as a secondary antibody.
Chromatin Immunoprecipitation Image 1: Phospho-Stat6 (Tyr641) (D8S9Y) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from Ramos cells, starved overnight then treated with IL-4 (100 ng/ml, 30 min) and either Phospho-Stat6 (Tyr641) (D8S9Y) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human MS4A1 Promoter Primers, SimpleChIP® Human DMD Intron 2 Primers #19133, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.