Recombinant Anti-STAT3 antibody [E121-21] (ab32500)
![Western blot - Anti-STAT3 antibody [E121-21] (ab32500)](https://www.abcam.com/ps/products/32/ab32500/Images/ab32500-478704-anti-stat3-antibody-e121-21-westernblot-human.jpg "Western blot - Anti-STAT3 antibody [E121-21] (ab32500)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E121-21] to STAT3
- Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-STAT3 antibody [E121-21]
See all STAT3 primary antibodies -
Description
Rabbit monoclonal [E121-21] to STAT3 -
Host species
Rabbit -
Specificity
The antibody only detects Stat3 without phosphorylation on Serine 727. It does not detect S727-phosphorylated Stat3. -
Tested applications
Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)more details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Human
Predicted to work with: Horse, Cow
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Immunogen
Synthetic peptide corresponding to Human STAT3. A synthetic peptide corresponding to residues surrounding Ser727 of human Stat3.
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Positive control
- HAP1, A431 cell lysate ICC/IF: HeLa cells Flow Cyt (intra): HeLa cells. IP: HeLa cells.
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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
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, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
E121-21 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Assay kits
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32500 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 |
|---|---|---|
| WB |
1/1000. Detects a band of approximately 92 kDa (predicted molecular weight: 88 kDa).
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| ICC/IF | (2) |
1/50.
For unpurified use at 1/250 dilution |
| IP |
1/20.
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| Flow Cyt (Intra) |
1/20.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/100 dilution |
| Notes |
|---|
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WB
1/1000. Detects a band of approximately 92 kDa (predicted molecular weight: 88 kDa). |
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ICC/IF
1/50. For unpurified use at 1/250 dilution |
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IP
1/20. |
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Flow Cyt (Intra)
1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/100 dilution |
Target
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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
modificationsTyrosine 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. - Information by UniProt
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Database links
- Entrez Gene: 508541 Cow
- Entrez Gene: 6774 Human
- Omim: 102582 Human
- SwissProt: P61635 Cow
- SwissProt: P40763 Human
- Unigene: 463059 Human
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Alternative names
- 1110034C02Rik antibody
- Acute Phase Response Factor antibody
- Acute-phase response factor antibody
see all
Images
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Western blot - Anti-STAT3 antibody [E121-21] (ab32500)All lanes : Anti-STAT3 antibody [E121-21] (ab32500) at 1/1000 dilution (Purified)
Lane 1 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate
Lane 2 : HaCaT (Human skin keratinocyte) whole cell lysate
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 88 kDa -
![Flow Cytometry (Intracellular) - Anti-STAT3 antibody [E121-21] (ab32500)](https://www.abcam.com/ps/products/32/ab32500/Images/ab32500-12-anti-stat3-antibody-e121-21-flowcytometry-a431-human.jpg)
Flow Cytometry (Intracellular) - Anti-STAT3 antibody [E121-21] (ab32500)Flow Cytometry analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labelling STAT3 with Purified ab32500 at 1:20 dilution (5 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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![Immunocytochemistry/ Immunofluorescence - Anti-STAT3 antibody [E121-21] (ab32500)](https://www.abcam.com/ps/products/32/ab32500/Images/ab32500-11-anti-stat3-antibody-e121-21-immunocytochemistry-shsy5y-human.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-STAT3 antibody [E121-21] (ab32500)Immunocytochemistry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling STAT3 with Purified ab32500 at 1:50 dilution (2.1 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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![Western blot - Anti-STAT3 antibody [E121-21] (ab32500)](https://www.abcam.com/ps/products/32/ab32500/Images/ab32500-277406-anti-stat3-antibody-e121-21-western-blot.jpg)
Western blot - Anti-STAT3 antibody [E121-21] (ab32500)Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: STAT3 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HEK293 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab32500 observed at 92 kDa. Red - loading control, ab8245, observed at 37 kDa.
Ab32500 detected the expected band for STAT3 in wild-type cells along with additional cross-reactive bands. The band was not seen in STAT3 knockout HAP1 cells. Wild-type and STAT3 knockout samples were subjected to SDS-PAGE. Ab32500 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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![Immunoprecipitation - Anti-STAT3 antibody [E121-21] (ab32500)](https://www.abcam.com/ps/products/32/ab32500/Images/ab32500-469039-antistat3antibodye12121immunoprecipitationhelahuman.jpg)
Immunoprecipitation - Anti-STAT3 antibody [E121-21] (ab32500)ab32500 at 1/20 dilution (0.5 µg) immunoprecipitating STAT3 in HeLa whole cell lysate.
Lane 1 (input): HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab32500 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32500 in HeLa whole cell lysateFor western blotting, ab32500 at 1/1000 and VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.
Blocking and diluting buffer: 5% NFDM /TBST.
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![Anti-STAT3 antibody [E121-21] (ab32500)](https://www.abcam.com/ps/products/32/ab32500/Images/ab32500-8-benefits-of-recombinant-antibodies.png)
Anti-STAT3 antibody [E121-21] (ab32500)
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
References (22)
ab32500 has been referenced in 22 publications.
- Li R et al. miR-21a inhibits decidual cell apoptosis by targeting Pdcd4. Genes Dis 8:171-180 (2021). PubMed: 33997164
- Wang C et al. Blocking the autocrine regulatory loop of Gankyrin/STAT3/CCL24/CCR3 impairs the progression and pazopanib resistance of clear cell renal cell carcinoma. Cell Death Dis 11:117 (2020). PubMed: 32051393
- Pu Z et al. Circular RNA circCUL3 Accelerates the Warburg Effect Progression of Gastric Cancer through Regulating the STAT3/HK2 Axis. Mol Ther Nucleic Acids 22:310-318 (2020). PubMed: 33230436
- Liu Z et al. STAT3-induced upregulation of long noncoding RNA HNF1A-AS1 promotes the progression of oral squamous cell carcinoma via activating Notch signaling pathway. Cancer Biol Ther 20:444-453 (2019). PubMed: 30404566
- Yang L et al. Interleukin-35 modulates the balance between viral specific CD4+CD25+CD127dim/- regulatory T cells and T helper 17 cells in chronic hepatitis B virus infection. Virol J 16:48 (2019). PubMed: 30992023