Revision 3

#4093

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, E-P

Reactivity:
H M R Mk

Sensitivity:
Endogenous

MW (kDa):
79, 86

Source/Isotype:
Rabbit IgG

UniProt ID:
#P40763

Entrez-Gene Id:
6774

Product Usage Information

Biotinylated antibodies are ideal for immunoassay technologies and high-throughput ELISA platforms that require antibody pairs where both antibodies are from the same host. Platforms utilizing biotinylated antibodies include, but are not limited to, MSD, xMAP, Quanterix Simoa, AlphaLISA, AlphaScreen, HTRF, LANCE, and TR-FRET.

Optimal dilutions/working concentrations should be determined by the end user. Please contact us if you require the antibody clone biotinylated at a different concentration, a carrier-free formulation, or a more customized packaging solution.

Storage

Supplied in 140 mM NaCl, 3 mM KCl, 10 mM sodium phosphate (pH 7.4) dibasic, 2 mM potassium phosphate monobasic, 2 mg/mL BSA, and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

Specificity/Sensitivity

Phospho-Stat3 (Tyr705) (D3A7) Rabbit Monoclonal Antibody (Biotinylated) detects endogenous levels of Stat3 only when phosphorylated at Tyr705. This antibody does not cross-react with phospho-EGFR or the corresponding phospho-tyrosines of other Stat proteins.

Species predicted to react based on 100% sequence homology

Hamster, Bovine, Pig, Horse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr705 of mouse Stat3.

Description

This Cell Signaling Technology® antibody is the biotinylated version of the unconjugated Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb #9145 and is expected to exhibit the same species cross-reactivity. The concentration of the biotinylated antibody is 500 ug/ml. Peptide ELISA data were generated using the biotinylated antibody. Western blot data were generated using the unconjugated antibody.

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).

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

W: Western Blotting E-P: Peptide ELISA (DELFIA)

Cross-Reactivity Key

H: Human M: Mouse R: Rat Mk: Monkey

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

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.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Western blot analysis of extracts from IFN-alpha treated Jurkat cells and HeLa cells (left), as well as EGF treated A431 cells (right), using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb. Note that the basal phospho-Stat3 in A431 is detected by the antibody.
Western Blotting Image 1: Phospho-Stat3 (Tyr705) (D3A7) Rabbit Monoclonal Antibody (Biotinylated)
Western blot analysis of extracts from HeLa cells, untreated or treated with IFNa (#36000, 100 ng/mL, 5 min) or IFNg (#80385, 100 ng/mL, 30 min); using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb #9145 (upper), Stat3 (D3Z2G) Rabbit mAb #12640 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 2: Phospho-Stat3 (Tyr705) (D3A7) Rabbit Monoclonal Antibody (Biotinylated)
Peptide ELISA was performed on a plate coated with a synthetic phosphopeptide corresponding to residues surrounding Tyr705 of mouse Stat3. The relationship between the concentration of Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb biotinylated (blue) and non-biotinylated (red) and the absorbance at 450 nm using a peptide-coated plate is shown. Streptavidin-HRP #3999, TMB Substrate #7004, and STOP Solution #7002 were used for color development.
E Image 1: Phospho-Stat3 (Tyr705) (D3A7) Rabbit Monoclonal Antibody (Biotinylated)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.