Name: Anti-STAT3 (phospho S727) antibody [E121-31]
Brand: RabMAb
SKU: ab32143
Rating: 5 (7 reviews)

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Immunocytochemistry/ Immunofluorescence - Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [E121-31] to STAT3 (phospho S727)
Suitable for: Flow Cyt, ELISA, ICC/IF, WB, Dot blot, IP, IHC-P
Reacts with: Mouse, Rat, Human

Carrier Free

Product name

Anti-STAT3 (phospho S727) antibody [E121-31]
See all STAT3 primary antibodies
Description

Rabbit monoclonal [E121-31] to STAT3 (phospho S727)
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt, ELISA, ICC/IF, WB, Dot blot, IP, IHC-Pmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Predicted to work with: Horse, Cow, Macaque monkey
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: A431 cell lysate, C6 treated with epidermal growth factor. IP: HeLa cells ICC/IF: A431 cells IHC-P: human astrocytoma, rat cerebral cortex, mouse liver, and brain astrocytoma tissues
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Mouse: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

E121-31
Isotype

IgG
Research areas

Alternative Versions
- Anti-STAT3 (phospho S727) antibody [E121-31] - BSA and Azide free (ab219593)
Assay kits
- STAT3 Transcription Factor Assay Kit (Colorimetric) (ab207229)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- A-431 whole cell lysate (ab7909)
Recombinant Protein
- Recombinant Human STAT3 protein (ab43618)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32143 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
Flow Cyt		Use at an assay dependent concentration.
ELISA		Use at an assay dependent concentration.
ICC/IF		1/500. For unpurified, use 1/100.
WB	(1)	1/1000 - 1/10000. Detects a band of approximately 98 kDa (predicted molecular weight: 88 kDa). Stimulation may be required to allow detection of the phosphorylated protein. Please see images below for recommended treatment conditions and positive controls.
Dot blot		1/1000.
IP		1/60.
IHC-P	(5)	1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified, use 1/50.

Notes
Flow Cyt Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
ICC/IF 1/500. For unpurified, use 1/100.
WB 1/1000 - 1/10000. Detects a band of approximately 98 kDa (predicted molecular weight: 88 kDa). Stimulation may be required to allow detection of the phosphorylated protein. Please see images below for recommended treatment conditions and positive controls.
Dot blot 1/1000.
IP 1/60.
IHC-P 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified, use 1/50.

Function

Signal transducer and transcription activator that mediates cellular responses to interleukins, KITLG/SCF, LEP and other growth factors. Once activated, recruits coactivators, such as NCOA1 or MED1, to the promoter region of the target gene (PubMed:17344214). May mediate cellular responses to activated FGFR1, FGFR2, FGFR3 and FGFR4. Binds to the interleukin-6 (IL-6)-responsive elements identified in the promoters of various acute-phase protein genes. Activated by IL31 through IL31RA. Involved in cell cycle regulation by inducing the expression of key genes for the progression from G1 to S phase, such as CCND1 (PubMed:17344214). Mediates the effects of LEP on melanocortin production, body energy homeostasis and lactation (By similarity). May play an apoptotic role by transctivating BIRC5 expression under LEP activation (PubMed:18242580). Cytoplasmic STAT3 represses macroautophagy by inhibiting EIF2AK2/PKR activity.
Tissue specificity

Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.
Involvement in disease

Hyperimmunoglobulin E recurrent infection syndrome, autosomal dominant
Autoimmune disease, multisystem, infantile-onset
Sequence similarities

Belongs to the transcription factor STAT family.
Contains 1 SH2 domain.
Post-translational
modifications

Tyrosine phosphorylated upon stimulation with EGF. Tyrosine phosphorylated in response to constitutively activated FGFR1, FGFR2, FGFR3 and FGFR4 (By similarity). Activated through tyrosine phosphorylation by BMX. Tyrosine phosphorylated in response to IL6, IL11, LIF, CNTF, KITLG/SCF, CSF1, EGF, PDGF, IFN-alpha, LEP and OSM. Activated KIT promotes phosphorylation on tyrosine residues and subsequent translocation to the nucleus. Phosphorylated on serine upon DNA damage, probably by ATM or ATR. Serine phosphorylation is important for the formation of stable DNA-binding STAT3 homodimers and maximal transcriptional activity. ARL2BP may participate in keeping the phosphorylated state of STAT3 within the nucleus. Upon LPS challenge, phosphorylated within the nucleus by IRAK1. Upon erythropoietin treatment, phosphorylated on Ser-727 by RPS6KA5. Phosphorylation at Tyr-705 by PTK6 or FER leads to an increase of its transcriptional activity. Dephosphorylation on tyrosine residues by PTPN2 negatively regulates IL6/interleukin-6 signaling.
Cellular localization

Cytoplasm. Nucleus. Shuttles between the nucleus and the cytoplasm. Translocated into the nucleus upon tyrosine phosphorylation and dimerization, in response to signaling by activated FGFR1, FGFR2, FGFR3 or FGFR4. Constitutive nuclear presence is independent of tyrosine phosphorylation. Predominantly present in the cytoplasm without stimuli. Upon leukemia inhibitory factor (LIF) stimulation, accumulates in the nucleus. The complex composed of BART and ARL2 plays an important role in the nuclear translocation and retention of STAT3. Identified in a complex with LYN and PAG1.
Target information above from: UniProt accession P40763 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 508541 Cow
- Entrez Gene: 6774 Human
- Entrez Gene: 20848 Mouse
- Entrez Gene: 25125 Rat
- Omim: 102582 Human
- SwissProt: P61635 Cow
- SwissProt: P40763 Human
- SwissProt: P42227 Mouse
- SwissProt: P52631 Rat
- Unigene: 463059 Human
- Unigene: 249934 Mouse
- Unigene: 473190 Mouse
- Unigene: 10247 Rat
see all
Alternative names
- 1110034C02Rik antibody
- Acute Phase Response Factor antibody
- Acute-phase response factor antibody
- ADMIO antibody
- APRF antibody
- AW109958 antibody
- DNA binding protein APRF antibody
- FLJ20882 antibody
- HIES antibody
- MGC16063 antibody
- Signal transducer and activator of transcription 3 (acute phase response factor) antibody
- Signal transducer and activator of transcription 3 antibody
- STAT 3 antibody
- Stat3 antibody
- STAT3_HUMAN antibody
see all

Immunocytochemistry/ Immunofluorescence - Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)

Purified ab32143 staining STAT3 (phospho S727) in A431 cells by Immunocytochemistry/ Immunofluorescence. 4% PFA-fixed, 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma) cells labelled with ab32143 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor 488) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing nuclear staining on A431 cell line. The red staining is ab7291 anti-Tubulin (mouse mAb), followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)

Immunohistochemical staining of paraffin embedded human astrocytoma with purified ab32143 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Western blot - Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)

All lanes : Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143) at 1/5000 dilution (purified)

Lane 1 : C6 treated with epidermal growth factor
Lane 2 : untreated C6 whole cell lysates

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 88 kDa
Observed band size: 98 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Western blot - Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)

All lanes : purified

Lane 1 : A431 treated with epidermal growth factor
Lane 2 : untreated A431 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 88 kDa
Observed band size: 98 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)

Immunohistochemical staining of paraffin embedded rat cerebral cortex with purified ab32143 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)

Immunohistochemical staining of paraffin embedded mouse liver with purified ab32143 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunocytochemistry/ Immunofluorescence - Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)

Immunocytochemical/Immunofluorescence analysis of untreated, EGF treated and EGF + LP treated A431 cells labelling STAT3 (phospho S727) with ab32143 (left) and STAT3 with ab32500 (right) both at a dilution of 1/500.

Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) (1/1000) was used as the secondary antibody (green). DAPI (blue) was used as the nuclear counterstain. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (1/200) was used as a counterstain (red).

The green staining was increased and translocated from the cytoplasm into the nucleus in the EGF (ab9697 100ng/ml, 10min) treated A431 cells when compared with A431 cells without treatment. After LP treatment, the green signal was decreased. For the pan antibody, there was no great difference after EGF (100ng/ml, 10min) or EGF (100ng/ml, 10min) + LP treatment.
Dot Blot - Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)

Dot Blot analysis of Lane 1: STAT3 (pS727) phospho peptide and Lane 2: STAT3 non-phospho peptide labeling STAT3 (phospho S727) with ab32143 at 1/1000 dilution (0.009 μg/ml). 5% NFDM /TBST was used as the diluting and blocking buffer and concentration. ab97051, Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100,000 dilution. Exposure time: 10 seconds.
Western blot - Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)

All lanes : Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143) at 1/1000 dilution (unpurified)

Lane 1 : A431 cell lysate
Lane 2 : A431 + EGF cell lysate

Predicted band size: 88 kDa
Observed band size: 98 kDa why is the actual band size different from the predicted?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)

IHC-P analysis of brain astrocytoma using unpurified ab32143 at 1/50 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunoprecipitation - Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)

ab32143 (purified) at 1/60 dilution (2.594 µg/ml) immunoprecipitating STAT3 in HeLa whole cell lysate.

Lane 1 (input): HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab32143 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32143 in HeLa whole cell lysate

For western blotting, ab32143 at 1/500 and VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/1000 dilution.

Blocking and diluting buffer: 5% NFDM /TBST.
Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab32143? Please let us know so that we can cite the reference in this datasheet.

ab32143 has been referenced in 55 publications.

Xu XS et al. SCARF1 promotes M2 polarization of Kupffer cells via calcium-dependent PI3K-AKT-STAT3 signalling to improve liver transplantation. Cell Prolif 54:e13022 (2021). PubMed: 33686740
Cui B et al. PKD3 promotes metastasis and growth of oral squamous cell carcinoma through positive feedback regulation with PD-L1 and activation of ERK-STAT1/3-EMT signalling. Int J Oral Sci 13:8 (2021). PubMed: 33692335
Yang W et al. Sinomenine Suppresses Development of Hepatocellular Carcinoma Cells via Inhibiting MARCH1 and AMPK/STAT3 Signaling Pathway. Front Mol Biosci 8:684262 (2021). PubMed: 34179090
Zhou Y et al. MiR-769-5p inhibits cancer progression in oral squamous cell carcinoma by directly targeting JAK1/STAT3 pathway. Neoplasma 67:528-536 (2020). PubMed: 32064884
Sun Z et al. NHE1 Mediates 5-Fu Resistance in Gastric Cancer via STAT3 Signaling Pathway. Onco Targets Ther 13:8521-8532 (2020). PubMed: 32904684

View all Publications for this product

Customer reviews and Q&As

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1-7 of 7 Abreviews or Q&A

Application

Immunohistochemistry (Frozen sections)

Sample

Mouse Tissue sections (tumor)

Permeabilization

Specification

tumor

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 31 2018

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (tumor xenograft)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: TE pH9

Specification

tumor xenograft

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Fixative

10% normal buffered formalin

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 12 2018

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Pig Tissue sections (lung)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: TE pH9

Permeabilization

Specification

lung

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Fixative

10% normal buffered formalin

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 12 2018

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (stomach)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: TE pH9

Permeabilization

Specification

stomach

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Fixative

10% normal buffered formalin

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 12 2018

Application

Western blot

Sample

Mouse Cell lysate - whole cell (pancreatic fiborblasts)

Gel Running Conditions

Reduced Denaturing (10% PAGE)

Loading amount

20 µg

Specification

pancreatic fiborblasts

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 01 2017

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (mouse pancreas)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: citrate

Permeabilization

Specification

mouse pancreas

Blocking step

casein as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.25% · Temperature: 4°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 01 2017

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Esophageal cancer)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: 10mM Tris - 1mM EDTA - 0.05% Tween-20, pH 9.0

Permeabilization

Specification

Esophageal cancer

Blocking step

(agent) for 20 minute(s) · Concentration: 100% · Temperature: 20°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Tatiana Karakasheva

Verified customer

Submitted Mar 04 2016

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Assay kits

Compatible Secondaries

Isotype control

Positive Controls

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (55)

Customer reviews and Q&As

Immunohistochemistry (Frozen sections) abreview for Anti-STAT3 (phospho S727) antibody [E121-31]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-STAT3 (phospho S727) antibody [E121-31]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-STAT3 (phospho S727) antibody [E121-31]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-STAT3 (phospho S727) antibody [E121-31]

Western blot abreview for Anti-STAT3 (phospho S727) antibody [E121-31]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-STAT3 (phospho S727) antibody [E121-31]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-STAT3 (phospho S727) antibody [E121-31]

Post-translational
modifications