Anti-STAT4 antibody [EPR25128-48] (ab284408)

Rabbit Recombinant Monoclonal STAT4 antibody. Suitable for IP, WB, IHC-P and reacts with Mouse, Rat, Human samples.

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Images

Western blot - Anti-STAT4 antibody [EPR25128-48] (AB284408), expandable thumbnail

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.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	IP	WB	IHC-P	Flow Cyt (Intra)
Human	Not recommended	Expected	Not recommended	Tested	Not recommended
Mouse	Not recommended	Tested	Tested	Tested	Not recommended
Rat	Not recommended	Tested	Tested	Tested	Not recommended

Not recommended
Not recommended

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

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/30	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/30	Notes -

Expected
Expected

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

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Human, Rat	Dilution info -	Notes -

Target data

See full target information STAT4

Function

Transcriptional regulator mainly expressed in hematopoietic cells that plays a critical role in cellular growth, differentiation and immune response (PubMed:10961885, PubMed:37256972, PubMed:8943379). Plays a key role in the differentiation of T-helper 1 cells and the production of interferon-gamma (PubMed:12213961, PubMed:35614130). Participates also in multiple neutrophil functions including chemotaxis and production of the neutrophil extracellular traps (By similarity). After IL12 binding to its receptor IL12RB2, STAT4 interacts with the intracellular domain of IL12RB2 and becomes tyrosine phosphorylated (PubMed:10415122, PubMed:7638186). Phosphorylated STAT4 then homodimerizes and migrates to the nucleus where it can recognize STAT target sequences present in IL12 responsive genes. Although IL12 appears to be the predominant activating signal, STAT4 can also be phosphorylated and activated in response to IFN-gamma stimulation via JAK1 and TYK2 and in response to different interleukins including IL23, IL2 and IL35 (PubMed:11114383, PubMed:34508746). Transcription activation of IFN-gamma gene is mediated by interaction with JUN that forms a complex that efficiently interacts with the AP-1-related sequence of the IFN-gamma promoter (By similarity). In response to IFN-alpha/beta signaling, acts as a transcriptional repressor and suppresses IL5 and IL13 mRNA expression during response to T-cell receptor (TCR) activation (PubMed:26990433).

Alternative names

Signal transducer and activator of transcription 4, STAT4

Recommended products

Rabbit Recombinant Monoclonal STAT4 antibody. Suitable for IP, WB, IHC-P and reacts with Mouse, Rat, Human samples.

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: EPR25128-48
Purification technique: Affinity purification Protein A
Specificity: Human species is recommended based on IHC-P results, we do not guarantee WB and IP for human.
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

The Signal Transducer and Activator of Transcription 4 (STAT4) is a critical member of the STAT protein family. It weighs approximately 85 kDa. STAT4 primarily functions as a transcription factor activated by phosphorylation through Janus kinases (JAKs) in response to cytokine signaling. This protein facilitates the translocation into the nucleus where it promotes the transcription of specific genes. STAT4 expression occurs mainly in T cells and natural killer (NK) cells indicating its role in the immune response.

Biological function summary

STAT4 is integral to the differentiation of naïve T cells into T helper 1 (Th1) cells. These Th1 cells play a central role in orchestrating immune responses against intracellular pathogens. STAT4 activation requires two distinct steps involving JAK kinases and subsequent binding to DNA regulatory elements of target genes. STAT4 does not function as part of a large protein complex but rather operates as a dimer following phosphorylation.

Pathways

STAT4 participates strongly in the JAK-STAT signaling pathway fundamental for immune cell maturation and response. The JAK-STAT pathway regulates several biological processes including proliferation and inflammation. STAT4 works closely with STAT1 in promoting Th1 differentiation while also interacting with other proteins like IL-12 to modulate immune activity. These interactions emphasize its influence in immune responses and could affect inflammatory pathways.

Associated diseases and disorders

STAT4 exhibits associations with autoimmune diseases such as rheumatoid arthritis and systemic lupus erythematosus. Aberrant STAT4 activity can lead to dysregulated immune responses making it a significant target in these disorders. In the context of rheumatoid arthritis altered STAT4 signaling enhances inflammation through its connection with other immune proteins like IL-23. Understanding STAT4's role in these diseases may offer avenues for therapeutic interventions and better management of immune-related conditions.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

16 product images

Western blot - Anti-STAT4 antibody [EPR25128-48] (ab284408)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Lane 1-3: This blot was developed using a higher sensitivity ECL substrate.
Lane 4: Lysates was made freshly and used in WB test immediately to minimize protein degradation.
The 37 kDa band in lane 1 would be a degradation band.
Negative control: NIH/3T3, EL4 (PMID: 8007943, 10358173)
Fresh lysates are preferred in this product.
Exposure time: Lane 1-3: 3 minutesLane 4: 26 seconds
All lanes: Western blot - Anti-STAT4 antibody [EPR25128-48] (ab284408) at 1/1000 dilution
Lane 1: F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 3: EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg
Lane 4: F9 (mouse embryonal carcinoma epithelial cell) whole cell fresh lysate at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 85 kDa
Observed band size: 81 kDa
Western blot - Anti-STAT4 antibody [EPR25128-48] (ab284408)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Negative control: brian, liver (PMID: 8007943)
The lower bands would be degradation bands.
Fresh lysates are preferred in this product.
Exposure time: 10 seconds
All lanes: Western blot - Anti-STAT4 antibody [EPR25128-48] (ab284408) at 1/1000 dilution
Lane 1: Mouse testis tissue lysate at 20 µg
Lane 2: Mouse brain tissue lysate at 20 µg
Lane 3: Mouse liver tissue lysate at 40 µg
Lane 4: Rat testis tissue lysate at 20 µg
Lane 5: Rat brain tissue lysate at 20 µg
Lane 6: Rat liver tissue lysate at 40 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 85 kDa
Observed band size: 81 kDa
Western blot - Anti-STAT4 antibody [EPR25128-48] (ab284408)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
The expression profile is consistent with what has been described in the literature (PMID: 8700209).
The lower bands would be degradation bands.
Fresh lysates are preferred in this product.
Exposure time: 15 seconds
All lanes: Western blot - Anti-STAT4 antibody [EPR25128-48] (ab284408) at 1/1000 dilution
Lane 1: Mouse thymus tissue lysate at 20 µg
Lane 2: Mouse lymph node tissue lysate at 20 µg
Lane 3: Rat thymus tissue lysate at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 85 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT4 antibody [EPR25128-48] (ab284408)
Immunohistochemical analysis of paraffin-embedded Rat testis tissue labelling STAT4 with ab284408 at 1/500 (1.136 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Postive staining on rat testis. The section was incubated with ab284408 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunoprecipitation - Anti-STAT4 antibody [EPR25128-48] (ab284408)
STAT4 was immunoprecipitated from 0.35 mg F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate 10 ug with ab284408 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab284408 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution. Lane 1: F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate 10 ug
Lane 2: ab284408 IP in F9 whole cell lysate 10 ug
Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab284408 in F9 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes
The 37 kDa band in lane 2 would be a degradation band.
All lanes: Immunoprecipitation - Anti-STAT4 antibody [EPR25128-48] (ab284408)
Predicted band size: 85 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT4 antibody [EPR25128-48] (ab284408)
Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labelling STAT4 with ab284408 at 1/500 (1.136 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Postive staining on mouse spleen. The section was incubated with ab284408 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins
Immunoprecipitation - Anti-STAT4 antibody [EPR25128-48] (ab284408)
STAT4 was immunoprecipitated from 0.35 mg Rat thymus tissue lysate 10 ug with ab284408 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab284408 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution. Lane 1: Rat thymus tissue lysate 10 ug
Lane 2: ab284408 IP in Rat thymus tissue lysate
Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab284408 in rat thymus tissue lysate 10 ug
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes
The lower band in lane 2 would be a degradation band.
All lanes: Immunoprecipitation - Anti-STAT4 antibody [EPR25128-48] (ab284408)
Predicted band size: 85 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT4 antibody [EPR25128-48] (ab284408)
Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labelling STAT4 with ab284408 at 1/500 (1.136 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Postive staining on mouse testis. The section was incubated with ab284408 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT4 antibody [EPR25128-48] (ab284408)
Immunohistochemical analysis of paraffin-embedded Mouse pancreatic carcinoma tissue labelling STAT4 with ab284408 at 1/500 (1.136 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Postive staining on immune cells in mouse pancreatic carcinoma. The section was incubated with ab284408 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT4 antibody [EPR25128-48] (ab284408)
Immunohistochemical analysis of paraffin-embedded Human spleen tissue labelling STAT4 with ab284408 at 1/500 (1.136 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Postive staining on human spleen. The section was incubated with ab284408 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT4 antibody [EPR25128-48] (ab284408)
Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labelling STAT4 with ab284408 at 1/500 (1.136 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Postive staining on immune cells in human breast carcinoma. The section was incubated with ab284408 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT4 antibody [EPR25128-48] (ab284408)
Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labelling STAT4 with ab284408 at 1/500 (1.136 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Negative control: no staining on human cardiac muscle. The section was incubated with ab284408 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT4 antibody [EPR25128-48] (ab284408)
Immunohistochemical analysis of paraffin-embedded Human testis tissue labelling STAT4 with ab284408 at 1/500 (1.136 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Postive staining on human testis. The section was incubated with ab284408 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT4 antibody [EPR25128-48] (ab284408)
Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labelling STAT4 with ab284408 at 1/500 (1.136 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Negative control: no staining on mouse cardiac muscle. The section was incubated with ab284408 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™, Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins
Western blot - Anti-STAT4 antibody [EPR25128-48] (ab284408)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The antibody detects Human STAT4 (phospho Y693).
This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.
In Western blot, anti-STAT4 antibody (ab284408) staining at 1/1000 dilution.
In Western blot, anti-His antibody (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.
In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
All lanes: Western blot - Anti-STAT4 (phospho Y693) antibody [EPR27081-49] (Anti-STAT4 (phospho Y693) antibody [EPR27081-49] ab313630) at 1/1000 dilution
Lane 1: 293T cells transfected with a Human STAT4 expression vector containing a His-tag were treated with treated with 100 mM Calyculin A for 30 minutes whole cell lysate at 20 µg
Lane 2: 293T cells transfected with a Human STAT4 (Y693F mutation) expression vector containing a His-tag were treated with 100 mM Calyculin A for 30 minutes whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Developed using the ECL technique.
Observed band size: 85 kDa
Exposure time: 180s
Western blot - Anti-STAT4 antibody [EPR25128-48] (ab284408)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.
In Western blot, anti-STAT4 antibody (ab284408) staining at 1/1000 dilution.
In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
All lanes: Western blot - Anti-STAT4 (phospho Y693) antibody [EPR27081-49] (Anti-STAT4 (phospho Y693) antibody [EPR27081-49] ab313630) at 1/1000 dilution
Lane 1: Untreated No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag) whole cell lysate (untreated membrane) at 20 µg
Lane 2: No-GFP-CD16.NK-92 treated with /mL IL-12 for 5 hours whole cell lysate (untreated membrane) at 20 µg
Lane 3: Untreated No-GFP-CD16.NK-92 whole cell lysate (phosphatase treated membrane) at 20 µg
Lane 4: No-GFP-CD16.NK-92 treated with /mL IL-12 for 5 hours whole cell lysate (phosphatase treated membrane) at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Developed using the ECL technique.
Observed band size: 85 kDa
Exposure time: 180s