Recombinant Anti-STAT5a antibody [E289] (ab32043)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E289] to STAT5a
- Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-STAT5a antibody [E289]
See all STAT5a primary antibodies -
Description
Rabbit monoclonal [E289] to STAT5a -
Host species
Rabbit -
Specificity
The antibody recognises Stat5a. It does not cross-react with other Stat family members. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details -
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: A431 cell lysate. IHC-P: Human squamous lung carcinoma. ICC/IF: Jurkat cells. Flow Cyt (intra): Jurkat 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.
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
E289 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32043 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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WB | (1) |
1/1000. Detects a band of approximately 92 kDa (predicted molecular weight: 91 kDa).
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IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. |
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ICC/IF |
1/100 - 1/500.
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IP |
1/20.
For unpurified use at 1/80. |
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Flow Cyt (Intra) |
1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.For unpurified use at 1/10. |
Notes |
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WB
1/1000. Detects a band of approximately 92 kDa (predicted molecular weight: 91 kDa). |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. |
ICC/IF
1/100 - 1/500. |
IP
1/20. For unpurified use at 1/80. |
Flow Cyt (Intra)
1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.For unpurified use at 1/10. |
Target
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Function
Carries out a dual function: signal transduction and activation of transcription. Binds to the GAS element and activates PRL-induced transcription. -
Sequence similarities
Belongs to the transcription factor STAT family.
Contains 1 SH2 domain. -
Post-translational
modificationsTyrosine phosphorylated in response to IL-2, IL-3, IL-7, IL-15, GM-CSF, growth hormone, prolactin, erythropoietin and thrombopoietin. Tyrosine phosphorylation is required for DNA-binding activity and dimerization. Serine phosphorylation is also required for maximal transcriptional activity. -
Cellular localization
Cytoplasm. Nucleus. Translocated into the nucleus in response to phosphorylation. - Information by UniProt
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Database links
- Entrez Gene: 6776 Human
- Entrez Gene: 20850 Mouse
- Entrez Gene: 24918 Rat
- Omim: 601511 Human
- SwissProt: P42229 Human
- SwissProt: P42230 Mouse
- SwissProt: Q62771 Rat
- Unigene: 437058 Human
see all -
Alternative names
- Mammary gland factor antibody
- MGF antibody
- Signal transducer and activator of transcription 5A antibody
see all
Images
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Anti-STAT5a antibody [E289] (ab32043) at 1/20000 dilution (purified) + K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 91 kDa
Observed band size: 92 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast tissue sections labeling STAT5a with purified ab32043 at 1:1000 dilution (0.12 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use). PBS instead of the primary antibody was used as the negative control.
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Immunocytochemistry/ Immunofluorescence analysis of Jurkat (human T cell leukemia T lymphocyte) cells labeling STAT5a with purified ab32043 at 1:100 (1.2 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 μg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Intracellular Flow Cytometry analysis of Jurkat (human acute T cell leukemia) cells labeling STAT5a with purified ab32043 at 1/100 dilution (1 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluorr® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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ab32043 (purified) at 1:20 dilution (0.6ug) immunoprecipitating in TF-1 whole cell lysate. TF-1 (Human Erythroleukemia erythroblast) whole cell lysate 10ug
Lane 2 (+): ab32043 & TF-1 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32043 in TF-1 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Unpurified ab32043 at 1/250 dilution, staining human squamous lung carcinoma by Immunohistochemistry, paraffin-embedded tissue. Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using (Tris/EDTA buffer, pH 9.0).
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All lanes : Anti-STAT5a antibody [E289] (ab32043) at 1/1000 dilution (Purified)
Lane 1 : Mouse brain lysates
Lane 2 : Rat brain lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 91 kDa
Observed band size: 92 kDa why is the actual band size different from the predicted? -
Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling STAT5a with unpurified ab32043 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.
Control: PBS only.
Nuclear counter stain: DAPI. -
Anti-STAT5a antibody [E289] (ab32043) at 1/1000 dilution (unpurified) + A431 cell lysate
Predicted band size: 91 kDa
Observed band size: 92 kDa why is the actual band size different from the predicted? -
Anti-STAT5a antibody [E289] (ab32043) at 1/1000 dilution (unpurified) +
Recombinant Human STAT5a protein (ab84627) at 0.01 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 91 kDa
Exposure time: 20 seconds -
Overlay histogram showing Jurkat cells stained with unpurified ab32043 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween 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 (ab32043, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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Tissue Microarrays stained for "Anti-STAT5a antibody [E289]” using "ab32043" in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab32043 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
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 (33)
ab32043 has been referenced in 33 publications.
- Yang Y et al. Targeting the STAT5A/IDO1 axis overcomes radioresistance and reverses the immunosuppressive tumor microenvironment in NSCLC. Int J Oncol 62:N/A (2023). PubMed: 36453241
- Wu S et al. Interplay between hypertriglyceridemia and acute promyelocytic leukemia mediated by the cooperation of peroxisome proliferator-activated receptor-α with the PML/RAR α fusion protein on super-enhancers. Haematologica 107:2589-2600 (2022). PubMed: 35546300
- Bailey JL et al. Oncostatin M Induces Lipolysis and Suppresses Insulin Response in 3T3-L1 Adipocytes. Int J Mol Sci 23:N/A (2022). PubMed: 35563078
- Xu S et al. Leukemia inhibitory factor is a therapeutic target for renal interstitial fibrosis. EBioMedicine 86:104312 (2022). PubMed: 36335669
- Jiang Y et al. Loss of STAT5A promotes glucose metabolism and tumor growth through miRNA-23a-AKT signaling in hepatocellular carcinoma. Mol Oncol 15:710-724 (2021). PubMed: 33155364