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 - ChIP Grade
- Suitable for: IHC-Fr, WB, IHC-P, IP, Flow Cyt, ChIP, ICC/IF
- Knockout validated
- Reacts with: Human
Product nameAnti-STAT3 antibody [E121-21] - ChIP Grade
See all STAT3 primary antibodies
DescriptionRabbit monoclonal [E121-21] to STAT3 - ChIP Grade
SpecificityThe antibody only detects Stat3 without phosphorylation on Serine 727. It does not detect S727-phosphorylated Stat3.
Tested applicationsSuitable for: IHC-Fr, WB, IHC-P, IP, Flow Cyt, ChIP, ICC/IFmore details
Species reactivityReacts with: Human
Predicted to work with: Horse, Cow
Synthetic peptide corresponding to Human STAT3. A synthetic peptide corresponding to residues surrounding Ser727 of human Stat3.
- HAP1, A431 cell lysate and lung squamous carcinoma. ICC/IF: HeLa cells Flow Cyt: HeLa cells. IP: HeLa cells
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
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.
|IHC-Fr||Use at an assay dependent concentration.|
|WB||1/1000. Detects a band of approximately 92 kDa (predicted molecular weight: 88 kDa).|
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|ChIP||Use at an assay dependent concentration.|
FunctionSignal 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 specificityHeart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.
Involvement in diseaseHyperimmunoglobulin E recurrent infection syndrome, autosomal dominant
Autoimmune disease, multisystem, infantile-onset
Sequence similaritiesBelongs to the transcription factor STAT family.
Contains 1 SH2 domain.
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 localizationCytoplasm. 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
- 1110034C02Rik antibody
- Acute Phase Response Factor antibody
- Acute-phase response factor antibody
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.
Immunohistochemical analysis of paraffin-embedded lung squamous carcinoma using ab32500 at a 1/50 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ab32500 staining STAT3 in HeLa cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab32500 at 1/250 dilution (shown in green) and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594), at 2µg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Chromatin was prepared from HepG2 + IL-6 (100 ng/ml 30 minutes) cells according to the Abcam X-ChIP protocol*. Cells were fixed with formaldehyde for 10 minutes.
The ChIP was performed with 25 µg of chromatin, 2 µg of ab32500 (red), and 20 µl of Protein A/G sepharose beads. 2 µg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
ab32500 (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 (+): ab32500 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32500 in HeLa whole cell lysate
For western blotting, ab32500 at 1/500 and VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM /TBST.
Overlay histogram showing HeLa cells stained with ab32500 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween 20 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32500, 1/100) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Anti-STAT3 antibody [E121-21] - ChIP Grade (ab32500) at 1/1000 dilution + A431 cell lysate
Predicted band size: 88 kDa
Observed band size: 92 kDa why is the actual band size different from the predicted?
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.
ab32500 has been referenced in 18 publications.
- 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
- Wu J et al. Impact of miR-223-3p and miR-2909 on inflammatory factors IL-6, IL-1ß, and TNF-a, and the TLR4/TLR2/NF-?B/STAT3 signaling pathway induced by lipopolysaccharide in human adipose stem cells. PLoS One 14:e0212063 (2019). PubMed: 30807577
- Wang CF et al. Knockdown of MSI1 inhibits the proliferation of human oral squamous cell carcinoma by inactivating STAT3 signaling. Int J Mol Med 44:115-124 (2019). PubMed: 31059073
- 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
- Wang C et al. Blocking the Feedback Loop between Neuroendocrine Differentiation and Macrophages Improves the Therapeutic Effects of Enzalutamide (MDV3100) on Prostate Cancer. Clin Cancer Res 24:708-723 (2018). PubMed: 29191973