Overview

  • Product name
    Anti-STAT5a antibody [E289] - BSA and Azide free
    See all STAT5a primary antibodies
  • Description
    Rabbit monoclonal [E289] to STAT5a - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide 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

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®).

    ab213219 is a PBS-only buffer formulated version of ab32043, containing no BSA or sodium azide, ideal for antibody labeling. Please refer to ab32043 for information on protocols, dilutions, and image data.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    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 rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab213219 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 Use at an assay dependent concentration. Detects a band of approximately 92 kDa (predicted molecular weight: 91 kDa).
IHC-P Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP Use at an assay dependent concentration.

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

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32043).

  • 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32043).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32043).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32043).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32043).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32043).

  • This IHC data was generated using the same anti-STAT5a antibody clone, E289, in a different buffer formulation (cat# ab32043).

    Unpurified ab32043 at 1/250 dilution, staining human squamous lung carcinoma by Immunohistochemistry, paraffin-embedded tissue.

References

This product has been referenced in:
  • Surana R  et al. IL4 limits the efficacy of tumor-targeted antibody therapy in a murine model. Cancer Immunol Res 2:1103-12 (2014). Read more (PubMed: 25204776) »
  • Cho KI  et al. Differential loss of prolyl isomerase or chaperone activity of Ran-binding protein 2 (Ranbp2) unveils distinct physiological roles of its cyclophilin domain in proteostasis. J Biol Chem 289:4600-25 (2014). WB ; Human . Read more (PubMed: 24403063) »
See all 6 Publications for this product

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