Recombinant Anti-STAT3 antibody [EPR787Y] (ab68153)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR787Y] to STAT3
- Suitable for: Flow Cyt (Intra), ChIC/CUT&RUN-seq, ICC/IF, WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-STAT3 antibody [EPR787Y]
See all STAT3 primary antibodies -
Description
Rabbit monoclonal [EPR787Y] to STAT3 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ChIC/CUT&RUN-seq, ICC/IF, WB, IHC-Pmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Rat and mouse heart tissue lysates; Human, mouse, and rat brain tissue lysates; Rat kidney tissue lysate; HeLa, HAP1, A431, HaCaT, NIH/3T3, C2C12, and Raji cell lysates. IHC-P: Human brain tissue. ICC/IF: HeLa cells. Flow Cyt (intra): Raji cells. ChIC/CUT&RUN seq: HepG2 cell
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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.
Properties
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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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR787Y -
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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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab68153 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 |
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Flow Cyt (Intra) |
1/30 - 1/50.
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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
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ICC/IF | (1) |
1/200.
For unpurified use at 1/140. |
WB | (7) |
1/1000 - 1/2000. Detects a band of approximately 75, 88 kDa (predicted molecular weight: 88 kDa).
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IHC-P | (1) |
1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
For unpurified use at 1/140. |
Notes |
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Flow Cyt (Intra)
1/30 - 1/50. |
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. |
ICC/IF
1/200. For unpurified use at 1/140. |
WB
1/1000 - 1/2000. Detects a band of approximately 75, 88 kDa (predicted molecular weight: 88 kDa). |
IHC-P
1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. For unpurified use at 1/140. |
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: 6774 Human
- Entrez Gene: 20848 Mouse
- Entrez Gene: 25125 Rat
- Omim: 102582 Human
- SwissProt: P40763 Human
- SwissProt: P42227 Mouse
- SwissProt: P52631 Rat
- Unigene: 463059 Human
see all -
Alternative names
- 1110034C02Rik antibody
- Acute Phase Response Factor antibody
- Acute-phase response factor antibody
see all
Images
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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/µL, 2.5 x 10^5 HepG2 cells (starved overnight and treated with 100ng/ml IL-6 for 30min) and 5 µg of ab68153 [EPR787Y]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
Additional screenshots of mapped reads can be downloaded here. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods. -
All lanes : Anti-STAT3 antibody [EPR787Y] (ab68153) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : STAT3 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 88 kDa
Observed band size: 92 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab68153 observed at 92 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab68153 was shown to react with STAT3 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255436 (knockout cell lysate ab263797) was used. Wild-type HeLa and STAT3 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab68153 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 antibody [EPR787Y] (ab68153)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human brain tissue sections labelling STAT3 with purified ab68153 at 1/200. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.
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All lanes : Anti-STAT3 antibody [EPR787Y] (ab68153) at 1/1000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HaCaT (Human skin keratinocyte) whole cell lysate
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 4 : C2C12 (Mouse myoblast cell line) whole cell lysate
Lane 5 : Human brain lysate
Lane 6 : Mouse brain lysate
Lane 7 : Rat brain lysate
Lane 8 : Rat kidney lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 88 kDa
Observed band size: 92 kDa why is the actual band size different from the predicted?
Exposure time: 90 secondsBlocking/Diluting buffer: 5% NFDM/TBST.
Rabbit monoclonal [EPR16891] to GAPDH (ab181602) used as loading control.
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All lanes : Anti-STAT3 antibody [EPR787Y] (ab68153) at 1/500 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : STAT3 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : HEK293 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 88 kDaLanes 1 - 4: Merged signal (red and green). Green - ab68153 observed at 92 kDa. Red - loading control, ab8245, observed at 37 kDa.
Ab68153 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. Ab68153 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/500 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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Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling STAT3 (green) with purified ab68153 at 1/200. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
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ab68153 stainingSTAT3in the human cell line HeLa (human cervix adenocarcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilized with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/30. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.
Isoytype control: Rabbit monoclonal IgG (Black)
Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (285)
ab68153 has been referenced in 285 publications.
- Liu A et al. Trimethylamine N-oxide promotes atherosclerosis via regulating the enriched abundant transcript 1/miR-370-3p/signal transducer and activator of transcription 3/flavin-containing monooxygenase-3 axis. Bioengineered 13:1541-1553 (2022). PubMed: 34923910
- Mei J et al. MACC1 regulates the AKT/STAT3 signaling pathway to induce migration, invasion, cancer stemness, and suppress apoptosis in cervical cancer cells. Bioengineered 13:61-70 (2022). PubMed: 34939526
- Shen J et al. Homeodomain-containing gene 10 contributed to breast cancer malignant behaviors by activating Interleukin-6/Janus kinase 2/Signal transducer and activator of transcription 3 pathway. Bioengineered 13:1335-1345 (2022). PubMed: 34983296
- Xue Y et al. Interferon-induced protein with tetratricopeptide repeats 1 (IFIT1) accelerates osteoclast formation by regulating signal transducer and activator of transcription 3 (STAT3) signalling. Bioengineered 13:2285-2295 (2022). PubMed: 35034537
- Luo Z et al. Construction and investigation of β3GNT2-associated regulatory network in esophageal carcinoma. Cell Mol Biol Lett 27:8 (2022). PubMed: 35073841