Name: Anti-STAT1 (phospho S727) antibody [EPR3146]
SKU: ab109461
Rating: 4 (1 reviews)

Rabbit Recombinant Monoclonal STAT1 phospho S727 antibody. Suitable for ChIC/CUT&RUN-seq, Dot, WB, IHC-P and reacts with Human, Rat, Mouse samples. Cited in 47 publications.

Images

Western blot - Anti-STAT1 (phospho S727) antibody [EPR3146] (AB109461), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ChIC/CUT&RUN-seq	Dot	WB	ICC/IF	IHC-P
Human	Tested	Expected	Tested	Not recommended	Tested
Mouse	Expected	Expected	Expected	Not recommended	Tested
Rat	Expected	Tested	Expected	Not recommended	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 5 µg	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat, Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Human, Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000 - 1/10000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Rat, Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes -
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/100 - 1/250	Notes Heat up to 98 °C, below boiling, and then let cool for 10-20 minutes. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/100 - 1/250	Notes Heat up to 98 °C, below boiling, and then let cool for 10-20 minutes. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/100 - 1/250	Notes Heat up to 98 °C, below boiling, and then let cool for 10-20 minutes. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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Target data

See full target information STAT1 phospho S727

Function

Signal transducer and transcription activator that mediates cellular responses to interferons (IFNs), cytokine KITLG/SCF and other cytokines and other growth factors (PubMed:12764129, PubMed:12855578, PubMed:15322115, PubMed:23940278, PubMed:34508746, PubMed:35568036, PubMed:9724754). Following type I IFN (IFN-alpha and IFN-beta) binding to cell surface receptors, signaling via protein kinases leads to activation of Jak kinases (TYK2 and JAK1) and to tyrosine phosphorylation of STAT1 and STAT2. The phosphorylated STATs dimerize and associate with ISGF3G/IRF-9 to form a complex termed ISGF3 transcription factor, that enters the nucleus (PubMed:28753426, PubMed:35568036). ISGF3 binds to the IFN stimulated response element (ISRE) to activate the transcription of IFN-stimulated genes (ISG), which drive the cell in an antiviral state (PubMed:28753426, PubMed:35568036). In response to type II IFN (IFN-gamma), STAT1 is tyrosine- and serine-phosphorylated (PubMed:26479788). It then forms a homodimer termed IFN-gamma-activated factor (GAF), migrates into the nucleus and binds to the IFN gamma activated sequence (GAS) to drive the expression of the target genes, inducing a cellular antiviral state (PubMed:8156998). Becomes activated in response to KITLG/SCF and KIT signaling (PubMed:15526160). May mediate cellular responses to activated FGFR1, FGFR2, FGFR3 and FGFR4 (PubMed:19088846). Following bacterial lipopolysaccharide (LPS)-induced TLR4 endocytosis, phosphorylated at Thr-749 by IKBKB which promotes binding of STAT1 to the 5'-TTTGAGGC-3' sequence in the ARID5A promoter, resulting in transcriptional activation of ARID5A and subsequent ARID5A-mediated stabilization of IL6 (PubMed:32209697). Phosphorylation at Thr-749 also promotes binding of STAT1 to the 5'-TTTGAGTC-3' sequence in the IL12B promoter and activation of IL12B transcription (PubMed:32209697). Involved in food tolerance in small intestine: associates with the Gasdermin-D, p13 cleavage product (13 kDa GSDMD) and promotes transcription of CIITA, inducing type 1 regulatory T (Tr1) cells in upper small intestine (By similarity).

Alternative names

Signal transducer and activator of transcription 1-alpha/beta, Transcription factor ISGF-3 components p91/p84, STAT1

Recommended products

Rabbit Recombinant Monoclonal STAT1 phospho S727 antibody. Suitable for ChIC/CUT&RUN-seq, Dot, WB, IHC-P and reacts with Human, Rat, Mouse samples. Cited in 47 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR3146
Purification technique: Affinity purification Protein A
Specificity: A phospho specific peptide corresponding to residues surrounding Serine 727 of human Stat-1 was used as an immunogen. This antibody only detects Stat-1 phosphorylated at Serine 727.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Signal Transducer and Activator of Transcription 1 (STAT1) is a protein of about 91 kDa commonly known simply as STAT1. It belongs to the STAT protein family which is involved in gene regulation. STAT1 expresses widely in many tissues including the immune system where it plays a major role. As an important player in cellular responses to cytokines and growth factors STAT1 becomes activated through phosphorylation often referred to as phospho-STAT1. This phosphorylation promotes dimerization an important step for its function.

Biological function summary

The STAT1 protein influences cell survival and immune responses. It functions as a transcription factor and participates in the interferon signaling pathway. In this setting it forms a complex with other STAT proteins such as STAT2 and STAT3 to regulate gene expression. By binding to specific DNA sequences in the nucleus STAT1 alters transcription in response to extracellular signals. It contributes to antiviral defense mechanisms and antiproliferative activities in various cells.

Pathways

The STAT1 protein plays essential roles in the JAK-STAT signaling pathway and interferon signaling pathway. It operates closely with proteins such as JAK1 and TYK2 which are involved in the phosphorylation process activating STAT1. During this activation STAT1 helps initiate cellular responses to interferons a type of signaling protein released during viral infections. The JAK-STAT pathway facilitates various physiological processes including immune responses and cell growth regulation.

Associated diseases and disorders

STAT1 dysregulation often connects to autoimmune diseases and infectious diseases. Elevated STAT1 activity associates with disorders like systemic lupus erythematosus where an imbalanced immune response occurs. Phosphorylation of STAT1 can also relate to chronic infections in cases where proper immune response activation becomes impaired. In certain cancers STAT1 interaction with proteins like SOCS1 might contribute to tumor immune evasion highlighting its vital role in pathology.

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10 product images

Western blot - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)
Blocking buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461) at 1/1000 dilution
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg
Lane 2: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate (treated with Alkaline Phosphatase for 1 hour) at 15 µg
Lane 3: Mouse brain lysate at 15 µg
Lane 4: Mouse brain lysate (treated with Alkaline Phosphatase for 1 hour) at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 87 kDa
Observed band size: 91 kDa
Western blot - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)
Blocking buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461) at 1/1000 dilution
Lane 1: Rat brain lysate at 15 µg
Lane 2: Rat brain lysate (treated with Alkaline Phosphatase for 1 hour) at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 87 kDa
Observed band size: 91 kDa
Western blot - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)
Blocking buffer: 2% BSA/TBST
Dilution buffer: 2% BSA/TBST
All lanes: Western blot - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461) at 1/5000 dilution
Lane 1: Untreated HeLa whole cell lysate at 10 µg
Lane 2: HeLa whole cell lysate treated with etoposide at 10 µg
Lane 3: HeLa whole cell lysate treated with etoposide, followed by membrane treatment with phosphatase at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 87 kDa
Observed band size: 90 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)
Immunohistochemical staining of paraffin embedded rat colon with purified ab109461 at a working dilution of 1/200. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 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-STAT1 (phospho S727) antibody [EPR3146] (ab109461)
Immunohistochemical staining of paraffin embedded mouse colon with purified ab109461 at a working dilution of 1/200. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 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-STAT1 (phospho S727) antibody [EPR3146] (ab109461)
Immunohistochemical staining of paraffin embedded human breast carcinoma with purified ab109461 at a working dilution of 1/200. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 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-STAT1 (phospho S727) antibody [EPR3146] (ab109461)
The lower section shows STAT1 detected with Anti-STAT1 alpha antibody [EPYR2154] ab92506, anti-STAT1 antibody, to confirm that the same amount of lysate is used in each lane.
Blocking and dilution biffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461) at 1/10000 dilution
Lane 1: Untreated HeLa (human cervix adenocarcinoma) at 10 µg
Lane 2: HeLa (human cervix adenocarcinoma) membrane treated with Alkaline Phosphatase at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1500 dilution
Developed using the ECL technique.
Predicted band size: 87 kDa
Observed band size: 91 kDa
Exposure time: 30s
Dot Blot - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)
Dot Blot analysis of Lane 1: STAT1 (pS727) phospho peptide and Lane 2: STAT1 non-phospho peptide, labeling STAT1 (phospho S727) with ab109461 at 1/1000 dilution. 5% NFDM/TBST was used as the blocking and diluting buffer. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody was used at 1/100000 dilution. Exposure time: 3 minutes.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)
Unpurified ab109461, at a 1/100 dilution, staining STAT1 (phospho S727) in paraffin embedded Human stomach adenocarcinoma tissue by Immunohistochemistry.
ChIC/CUT&RUN sequencing - Anti-STAT1 (phospho S727) antibody [EPR3146] (ab109461)
ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/µL, 2 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell line) treated with IFN gamma (50ng/ml 1h) cells and 5µg of ab109461 [EPR3146]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 is also shown.

Additional screenshots of mapped reads can be found in the Protocol booklet in the Support and downloads section.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.