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RabMAb

Anti-STAT5a 抗体 [E289] (ab32043)

製品の概要

  • 製品名
    Anti-STAT5a antibody [E289]
    STAT5a 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [E289] to STAT5a
  • 由来種
    Rabbit
  • 特異性
    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.
  • アプリケーション
    適用あり: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

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

  • ポジティブ・コントロール
    • WB: A431 cell lysate. IHC-P: Human squamous lung carcinoma. ICC/IF: Jurkat cells. Flow Cyt: Jurkat cells.
  • 特記事項

    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.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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. 

ターゲット情報

  • 機能
    Carries out a dual function: signal transduction and activation of transcription. Binds to the GAS element and activates PRL-induced transcription.
  • 配列類似性
    Belongs to the transcription factor STAT family.
    Contains 1 SH2 domain.
  • 翻訳後修飾
    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.
  • 細胞内局在
    Cytoplasm. Nucleus. Translocated into the nucleus in response to phosphorylation.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

画像

  • 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).

  • 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?)

  • 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.

参考文献

This product has been referenced in:
  • 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) »
  • Guo H  et al. Functional defects in CD4(+) CD25(high) FoxP3(+) regulatory cells in ankylosing spondylitis. Sci Rep 6:37559 (2016). Read more (PubMed: 27901054) »

See all 9 Publications for this product

レビューと Q&A

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
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投稿 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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