Overview

  • 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, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    within Human STAT5a aa 750 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P42229

  • Positive control
    • WB: A431 cell lysate. IHC-P: Human squamous lung carcinoma. ICC/IF: Jurkat cells. Flow Cyt: Jurkat cells.
  • General notes

    A trial size is available to purchase for this antibody.

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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
WB 1/1000. Detects a band of approximately 92 kDa (predicted molecular weight: 91 kDa).
IHC-P 1/1000.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
See IHC antigen retrieval protocols

ICC/IF 1/100 - 1/500.
Flow Cyt 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
For unpurified use at 1/10.

IP 1/20.

For unpurified use at 1/80. 

Target

  • 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
    modifications
    Tyrosine 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
  • Database links
  • Alternative names
    • Mammary gland factor antibody
    • MGF antibody
    • Signal transducer and activator of transcription 5A antibody
    • STA5A_HUMAN antibody
    • STAT 5 antibody
    • STAT 5A antibody
    • STAT5 antibody
    • STAT5A antibody
    see all

Images

  • 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.

  • 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.

  • 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 Fluor® 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).

  • 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 secondary antibody (HRP) (ab131366) was used as the secondary antibody 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.

  • 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 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. 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.

References

This product has been referenced in:
  • Wang S  et al. Exosomes released by hepatocarcinoma cells endow adipocytes with tumor-promoting properties. J Hematol Oncol 11:82 (2018). Read more (PubMed: 29898759) »
  • He L  et al. Local blockage of self-sustainable erythropoietin signaling suppresses tumor progression in non-small cell lung cancer. Oncotarget 8:82352-82365 (2017). Read more (PubMed: 29137269) »
See all 11 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
Application
ChIP
Sample
Human Cell lysate - whole cell (Human Breast Cancer --T47D)
Negative control
non-specific rabbit IgG
Specification
Human Breast Cancer --T47D
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 540 ul 37% Formaldehyde into 2
Positive control
+Prolactin 100ng for 45 minutes qRT-PCR looking at STAT5A recruitment to -700kb of known STAT5A target gene (CISH)

Abcam user community

Verified customer

Submitted Nov 02 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (macrophage)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
10 µg
Specification
macrophage
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 02 2016

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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