Recombinant Anti-STAT6 antibody [YE361] (ab32520)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [YE361] to STAT6
- Suitable for: ChIC/CUT&RUN-seq, Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-STAT6 antibody [YE361]
See all STAT6 primary antibodies -
Description
Rabbit monoclonal [YE361] to STAT6 -
Host species
Rabbit -
Specificity
This antibody does not cross-react with other Stat family members.
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Tested applications
Suitable for: ChIC/CUT&RUN-seq, Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide within Human STAT6 aa 800 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P42226 -
Positive control
- WB: Raji, NIH/3T3 and RAW 264.7 cell lysates. IHC-P: Human skin carcinoma, kidney, transitional cell carcinoma of bladder, glioma and solitary fibrous tumor tissues; Mouse kidney tissue. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells. IP: NIH/3T3 cell lysate. ChIC/CUT&RUN-Seq: 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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
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: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
YE361 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-STAT6 antibody [YE361] (ab196478)
- HRP Anti-STAT6 antibody [YE361] (ab196480)
- Alexa Fluor® 647 Anti-STAT6 antibody [YE361] (ab196928)
- Alexa Fluor® 594 Anti-STAT6 antibody [YE361] (ab207012)
- Alexa Fluor® 405 Anti-STAT6 antibody [YE361] (ab207013)
- Alexa Fluor® 555 Anti-STAT6 antibody [YE361] (ab207014)
- Alexa Fluor® 568 Anti-STAT6 antibody [YE361] (ab207015)
- Anti-STAT6 antibody [YE361] - BSA and Azide free (ab215995)
- PE Anti-STAT6 antibody [YE361] (ab223917)
- FITC Anti-STAT6 antibody [YE361] (ab223918)
- APC Anti-STAT6 antibody [YE361] (ab223919)
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Compatible Secondaries
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Isotype control
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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 ab32520 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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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
5µg |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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WB | (1) |
1/1000 - 1/2000. Detects a band of approximately 100 kDa (predicted molecular weight: 94 kDa).
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IHC-P |
1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ICC/IF |
1/100.
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IP |
1/20 - 1/80.
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Notes |
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ChIC/CUT&RUN-seq
Use at an assay dependent concentration. 5µg |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
1/1000 - 1/2000. Detects a band of approximately 100 kDa (predicted molecular weight: 94 kDa). |
IHC-P
1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
1/100. |
IP
1/20 - 1/80. |
Target
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Function
Carries out a dual function: signal transduction and activation of transcription. Involved in interleukin-4 signalling. -
Sequence similarities
Belongs to the transcription factor STAT family.
Contains 1 SH2 domain. -
Post-translational
modificationsTyrosine phosphorylated following stimulation by IL-4 and IL-3. -
Cellular localization
Cytoplasm. Nucleus. Translocated into the nucleus in response to phosphorylation. - Information by UniProt
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Database links
- Entrez Gene: 6778 Human
- Entrez Gene: 20852 Mouse
- Omim: 601512 Human
- SwissProt: P42226 Human
- SwissProt: P52633 Mouse
- Unigene: 524518 Human
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Alternative names
- 12S1644 antibody
- D12S1644 antibody
- IL 4 STAT antibody
see all
Images
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Immunohistochemical analysis of formalin fixed paraffin embedded human kidney labelling STAT6 with ab32520 at a concentration of 0.79µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 min with DISCOVERY cell conditioning solution (CC1) 100°C, pH 8.5. ab32520 was incubated at 37°C for 16 min. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell line) cells and 5µg of ab32520 [YE361]. 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.
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All lanes : Anti-STAT6 antibody [YE361] (ab32520) at 1/1000 dilution (purified)
Lane 1 : NIH/3T3 (mouse embyro fibroblast cell line) cell lysate
Lane 2 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution
Predicted band size: 94 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
Immunohistochemical staining of paraffin embedded human kidney with purified ab32520 at a working dilution of 1/50. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 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.
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Intracellular Flow Cytometry analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling STAT6 with purified ab32520 at 1/30 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.
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Immunofluorescence staining of HeLa (human epithelial cell line from cervix adenocarcinoma) cells with purified ab32520 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab32520 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
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ab32520 (purified) at 1/20 immunoprecipitating STAT6 in 10 μg NIH/3T3 (mouse embyro fibroblast cell line; Lanes 1 and 2, observed at 100 kDa). Lane 3 - PBS. For western blotting, HRP Veriblot for IP (ab131366) was used for detection (1/10 000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labelling STAT6 with unpurified ab32520 at a dilution of 1/1000. HRP goat anti-rabbit (ab97051) was used at a dilution of 1/500. The antigen retrieval solution was Tris-EDTA buffer, pH 9.0.
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Anti-STAT6 antibody [YE361] (ab32520) at 1/20000 dilution (purified) + Raji (human Burkitt's lymphoma cell line) cell lysate at 10 µg
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution
Predicted band size: 94 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skin carcinoma tissue labelling STAT6 with unpurified ab32520 at 1/100.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunocytochemistry/Immunofluorescence analyis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labelling STAT6 with unpurified ab32520 at 1/100.
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Anti-STAT6 antibody [YE361] (ab32520) at 1/1000 dilution (unpurified) + NIH/3T3 (mouse embyro fibroblast cell line) cell lysate
Predicted band size: 94 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human transitional cell carcinoma of bladder tissue sections labelling STAT6 with unpurified ab32520 at a dilution of 1/1000. HRP goat anti-rabbit (ab97051) was used at a dilution of 1/500. The antigen retrieval solution was Tris-EDTA buffer, pH 9.0.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human glioma tissue sections labelling STAT6 with unpurified ab32520 at a dilution of 1/1000. HRP goat anti-rabbit (ab97051) was used at a dilution of 1/500. The antigen retrieval solution was Tris-EDTA buffer, pH 9.0.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labelling STAT6 with unpurified ab32520 at a dilution of 1/1000. HRP goat anti-rabbit (ab97051) was used at a dilution of 1/500. The antigen retrieval solution was Tris-EDTA buffer, pH 9.0.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human solitary fibrous tumor tissue sections labelling STAT6 with unpurified ab32520 at a dilution of 1/1000. HRP goat anti-rabbit (ab97051) was used at a dilution of 1/500. The antigen retrieval solution was Tris-EDTA buffer, pH 9.0.
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 (60)
ab32520 has been referenced in 60 publications.
- Lv C et al. Narciclasine targets STAT3 via distinct mechanisms in tamoxifen-resistant breast cancer cells. Mol Ther Oncolytics 24:340-354 (2022). PubMed: 35118192
- Yu Y et al. Targeting Forkhead box O1-aquaporin 5 axis mitigates neuropathic pain in a CCI rat model through inhibiting astrocytic and microglial activation. Bioengineered 13:8567-8580 (2022). PubMed: 35324416
- Gao P et al. STAT6 Upregulates NRP1 Expression in Endothelial Cells and Promotes Angiogenesis. Front Oncol 12:823377 (2022). PubMed: 35600336
- Zou W et al. Conditioned medium from the bone marrow mesenchymal stem cells modulates immune response via signal transduction and activator of transcription 6 signaling pathway in an allergic rhinitis mouse model. Allergol Immunopathol (Madr) 50:105-114 (2022). PubMed: 35789409
- Glaubitz J et al. In mouse chronic pancreatitis CD25+FOXP3+ regulatory T cells control pancreatic fibrosis by suppression of the type 2 immune response. Nat Commun 13:4502 (2022). PubMed: 35922425