Revision 8
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, IHC-P, IF-IC, FC-FP

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

90

Source/Isotype:

Rabbit IgG

UniProt ID:

#P42229, #P51692

Entrez-Gene Id:

6776, 6777

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunohistochemistry (Paraffin) 1:200 - 1:800
Immunofluorescence (Immunocytochemistry) 1:100 - 1:400
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:400

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 #40284.

Specificity / Sensitivity

Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb detects endogenous levels of Stat5a only when phosphorylated at Tyr694 and Stat5b when phosphorylated at Tyr699.

Species Reactivity:

Human, Mouse

Species predicted to react based on 100% sequence homology

Rat, Bovine

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Tyr694 of Stat5.

Background

Stat5 is activated in response to a wide variety of ligands, including IL-2, GM-CSF, growth hormone, and prolactin. Phosphorylation at Tyr694 is obligatory for Stat5 activation (1,2). This phosphorylation is mediated by Src upon erythropoietin stimulation (3). Stat5 is constitutively active in some leukemic cell types (4). Phosphorylated Stat5 is found in some endothelial cells treated with IL-3, which suggests its involvement in angiogenesis and cell motility (5). Stat5a and Stat5b are independently regulated and activated in various cell types. For instance, interferon treatment predominantly activates Stat5a in U-937 cells and Stat5b in HeLa cells (6).

  1. Gouilleux, F. et al. (1994) EMBO J 13, 4361-9.
  2. Wakao, H. et al. (1994) EMBO J 13, 2182-91.
  3. Okutani, Y. et al. (2001) Oncogene 20, 6643-50.
  4. Demoulin, J.B. et al. (1999) J Biol Chem 274, 25855-61.
  5. Dentelli, P. et al. (1999) J Immunol 163, 2151-9.
  6. Meinke, A. et al. (1996) Mol Cell Biol 16, 6937-44.

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

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.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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 8
#9314

Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb

Western Blotting Image 1: Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb Expand Image
Western blot analysis of extracts from K562, untreated or treated with Gleevec for 1 hour, using Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb (upper) or Stat5 Antibody #9310 (lower).
Immunohistochemistry Image 1: Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb.
Immunohistochemistry Image 2: Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded K562 cell pellets control (left) or STI-571-treated (right) using Phospho-Stat5 (Tyr694) Rabbit mAb.
Immunohistochemistry Image 3: Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb.
Immunofluorescence Image 1: Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of HeLa cells, IFNα-treated (left) or untreated (right), using Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb (green) and Pan-Keratin (C11) Mouse mAb #4545 (red).
Flow Cytometry Image 1: Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb Expand Image
Flow cytometric analysis of TF-1 cells, untreated (blue) or treated with hGM-CSF #8922 (50 ng/mL, 15 min; green) using Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb (solid lines) or 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.