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Purified anti-STAT3 Phospho (Ser727) Antibody

Pricing & Availability
Clone
A16089A (See other available formats)
Regulatory Status
RUO
Other Names
Signal Transducer and Activator of Transcription 3, Acute-Phase Response Factor, APRF, ADMIO, HIES
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
A16089A_PURE_STAT3-Phospho-Ser727_Antibody_1_022221
Total cell lysates (15 µg total protein) from Jurkat cells serum starved and untreated (-) or stimulated with 160 nM PMA for 15 minutes (+) were resolved by 4-20% Tris-glycine gel electrophoresis, transferred to a nitrocellulose membrane, and probed with 2.0 µg/mL (1:250 dilution) purified anti-STAT3 Phospho (Ser727) antibody (clones A16089A and A16089B) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Purified anti-STAT3 pan antibody (clone 4G4B45) (Cat. No. 678002) was used as a pan STAT3 loading control at 1.0 µg/mL (1:500 dilution). Lane M: Molecular weight marker.
  • A16089A_PURE_STAT3-Phospho-Ser727_Antibody_1_022221
    Total cell lysates (15 µg total protein) from Jurkat cells serum starved and untreated (-) or stimulated with 160 nM PMA for 15 minutes (+) were resolved by 4-20% Tris-glycine gel electrophoresis, transferred to a nitrocellulose membrane, and probed with 2.0 µg/mL (1:250 dilution) purified anti-STAT3 Phospho (Ser727) antibody (clones A16089A and A16089B) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Purified anti-STAT3 pan antibody (clone 4G4B45) (Cat. No. 678002) was used as a pan STAT3 loading control at 1.0 µg/mL (1:500 dilution). Lane M: Molecular weight marker.
  • A16089A_PURE_STAT3-Phospho-Ser727_Antibody_2_022221
    Total cell lysates (15 µg total protein) from HeLa and NIH/3T3 cells serum starved and untreated (-) or stimulated with 100 ng/mL recombinant human IL-6 (Cat. No. 715104) or recombinant mouse IL-6 (Cat. No. 715202), respectively for 30 minutes (+) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL (1:2500 dilution) purified anti-STAT3 Phospho (Ser727) antibody (clones A16089A and A16089B) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG Antibody (Cat. No. 405306) at a 1:3000 dilution. Lane M: Molecular weight marker. Equal protein loading was confirmed using Direct-Blot™ HRP anti-GAPDH Antibody (Cat. No. 607904).
  • A16089A_PURE_STAT3-Phospho-Ser727_Antibody_3_022221
    Total cell lysates (15 µg total protein) from HeLa and cells serum starved and untreated (-) or stimulated with 100 ng/mL recombinant human IL-6 for 30 minutes were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL (1:2500 dilution) purified anti-STAT3 Phospho (Ser727) antibody (clones A16089A and A16089B) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. To demonstrate phospho-specificity of the clone, membranes were either untreated or pre-treated with lambda protein phosphatase. Equal protein loading was confirmed using Direct-Blot™ HRP anti-GAPDH Antibody (Cat. No. 607904). Lane M: Molecular weight marker.
  • A16089A_PURE_STAT3-Phospho-Ser727_Antibody_4_022221
    HeLa cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with Triton X-100 for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with a 1:200 dilution (2.5 µg/mL) of purified anti-STAT3 Phospho (Ser727) (clones A16089A and A16089B) for two hours at room temperature, followed by incubation with Alexa Fluor® 594 goat anti-mouse IgG (Cat. No. 405326) at 2.0 µg/mL. To confirm phospho-specificity of each clone, fixed cells pre-treated with lambda protein phosphatase (LPP) prior to staining were included as a control. Nuclei were counterstained with DAPI, and the image was captured with a 60X objective.
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669501 25 µg $112
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669502 100 µg $265
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Description

The STAT3 transcription factor is an important signaling molecule for many cytokines and growth factor receptors and is required for murine fetal development. STAT3 is a member of the STAT (signal transducer and activator of transcription) protein family that is phosphorylated in response to a cytokine receptor-associated kinase activity. STAT3 is activated by phosphorylation at Tyr705, which induces dimerization, nuclear translocation, and DNA binding. Transcriptional activation was reported to be regulated by phosphorylation at Ser727 through MAPK and mTOR pathways.

STAT3 forms both homo- and heterotrimers and is involved in the activation of genes required for cell growth and apoptosis. STAT3 is also involved in gp130 signaling and binds to IL-6 response elements in various acute phase protein promoters. STAT3 is phosphorylated by signaling from IFNs, EGF, FGF, IL-5, HGF, LIF, and BMP-2. STAT3 activity is inhibited by PIAS3 and GRIM-19 and can also be regulated by the Rac1 protein.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Synthetic human STAT3 peptide phosphorylated at serine 727
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 1.0 - 2.0 µg/mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

Clones A16089A and A16089B are both specific for STAT3 phosphorylated at serine 727, but performed differently for different applications tested:

Both clones displayed strong specificity and selectivity for WB, but clone A16089A displayed a considerably stronger affinity for the target compared to A16089B

Both clones performed well for ICC, though clone A16089A displayed mild extranuclear staining

A16089B performed well for ChIP, A16089A failed ChIP application testing

RRID
AB_2892483 (BioLegend Cat. No. 669501)
AB_2892483 (BioLegend Cat. No. 669502)

Antigen Details

Structure
STAT3 is a 770 amino acid protein with a predicted molecular weight of 88 kDa. It consists of a DNA binding domain, a SH2 domain, a regulatory tyrosine responsible for binding of SH2 domain, and a C-terminal transactivation domain.
Distribution

Cytosol and nucleus/Ubiquitously expressed

Function
Transcription factor
Antigen References
  1. Akira S, et al. 1994. Cell. 77:63-71.
  2. Zhang X, et al. 1995. Science. 267:1990-4.
  3. Sanchez-Margalet V & Martin-Romero C. 2001. Cell Immunol. 211:30-6.
  4. Simon AR, et al. 2000. Science. 290:144-7.
  5. Hoey T & Grusby MJ. 1999. Adv Immunol. 71:145-62.
Gene ID
6774 View all products for this Gene ID
UniProt
View information about STAT3 Phospho Ser727 on UniProt.org
Go To Top Version: 1    Revision Date: 02/22/2021

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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