Revision 4
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:

IP, IF-IC, FC-FP, ChIP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

86

Source/Isotype:

Rabbit IgG

UniProt ID:

#P40763

Entrez-Gene Id:

6774

Product Information

Product Usage Information

For optimal ChIP results, use 10 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
Immunoprecipitation 1:200
Immunofluorescence (Immunocytochemistry) 1:800
Flow Cytometry (Fixed/Permeabilized) 1:50
Chromatin IP 1:50

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 #50804.

Specificity / Sensitivity

Phospho-Stat3 (Ser727) (D4X3C) Rabbit mAb is recognizes endogenous levels of Stat3 protein only when phosphorylated at Ser727.

Species Reactivity:

Human

Species predicted to react based on 100% sequence homology

Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phospho-peptide corresponding to residues surrounding Ser727 of human Stat3 protein.

Background

The Stat3 transcription factor is an important signaling molecule for many cytokines and growth factor receptors (1) and is required for murine fetal development (2). Research studies have shown that Stat3 is constitutively activated in a number of human tumors (3,4) and possesses oncogenic potential (5) and anti-apoptotic activities (3). Stat3 is activated by phosphorylation at Tyr705, which induces dimerization, nuclear translocation, and DNA binding (6,7). Transcriptional activation seems to be regulated by phosphorylation at Ser727 through the MAPK or mTOR pathways (8,9). Stat3 isoform expression appears to reflect biological function as the relative expression levels of Stat3α (86 kDa) and Stat3β (79 kDa) depend on cell type, ligand exposure, or cell maturation stage (10). It is notable that Stat3β lacks the serine phosphorylation site within the carboxy-terminal transcriptional activation domain (8).

  1. Heim, M.H. (2001) J Recept Signal Transduct Res 19, 75-120.
  2. Takeda, K. et al. (1997) Proc Natl Acad Sci U S A 94, 3801-4.
  3. Catlett-Falcone, R. et al. (1999) Immunity 10, 105-15.
  4. Garcia, R. and Jove, R. (1998) J Biomed Sci 5, 79-85.
  5. Bromberg, J.F. et al. (1999) Cell 98, 295-303.
  6. Darnell, J.E. et al. (1994) Science 264, 1415-21.
  7. Ihle, J.N. (1995) Nature 377, 591-4.
  8. Wen, Z. et al. (1995) Cell 82, 241-50.
  9. Yokogami, K. et al. (2000) Curr Biol 10, 47-50.
  10. Biethahn, S. et al. (1999) Exp Hematol 27, 885-94.

Species Reactivity

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

Applications Key

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 4
#34911

Phospho-Stat3 (Ser727) (D4X3C) Rabbit mAb

Immunoprecipitation Image 1: Phospho-Stat3 (Ser727) (D4X3C) Rabbit mAb Expand Image
Immunoprecipitation of Phospho Stat3 (Ser727) from U266 cells treated with Human Interferon-α1 #93613 (hIFNa1; 50 ng/mL, 15 min). Lane 1 is 10% input, lane 2 is precipitated with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-Stat3 (Ser727) (D4X3C) Rabbit mAb, #34911. Western blot was performed using Phospho-Stat3 (Ser727) (D8C2Z) Rabbit mAb, #94994.
Immunoprecipitation Image 2: Phospho-Stat3 (Ser727) (D4X3C) Rabbit mAb Expand Image
Immunoprecipitation of Phospho Stat3 (Ser727) from A431 cells treated with Human Epidermal Growth Factor #72528 (hEGF; 100 ng/mL, 30 min). Lane 1 is 10% input, lane 2 is precipitated with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-Stat3 (Ser727) (D4X3C) Rabbit mAb, #34911. Western blot was performed using Phospho-Stat3 (Ser727) (D8C2Z) Rabbit mAb, #94994.
Immunofluorescence Image 1: Phospho-Stat3 (Ser727) (D4X3C) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of A-431 cells (Stat3-expressing), either serum-starved and treated with Wortmannin #9915 (1 uM) and U0126 #9903 (10 uM, 2 hours; left panel) or Human Epidermal Growth Factor (hEGF) #8916 (100 ng/mL, 30 minutes; center panel), and PC-3 cells serum-starved and treated with hEGF #8916 (right panel; Stat3-negative), using Phospho-Stat3 (Ser727) (D4X3C) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red; top panel only).
Flow Cytometry Image 1: Phospho-Stat3 (Ser727) (D4X3C) Rabbit mAb Expand Image
Flow cytometric analysis of human peripheral blood mononuclear cells, untreated (left) or TPA-treated #4174 (40nM, 15 minutes; right), using Phospho-Stat3 (Ser727) (D4X3C) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) was used as a secondary antibody.
Chromatin Immunoprecipitation Image 1: Phospho-Stat3 (Ser727) (D4X3C) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 Hep G2 cells starved overnight and treated with IL-6 (100ng/mL) for 30 minutes and either 10 μl of Phospho-Stat3 (Ser727) (D4X3C) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human c-Fos Promoter Primers #4663, human IRF-1 promoter primers, 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.