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    stat5a-antibody-e289-ab32043.pdf

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Signal Transduction Signaling Pathway Nuclear Signaling STATs
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RecombinantRabMAb

Recombinant Anti-STAT5a antibody [E289] (ab32043)

  • Datasheet
Reviews (2) Submit a question References (25)

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Western blot - Anti-STAT5a antibody [E289] (ab32043)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a antibody [E289] (ab32043)
  • Immunocytochemistry/ Immunofluorescence - Anti-STAT5a antibody [E289] (ab32043)
  • Flow Cytometry (Intracellular) - Anti-STAT5a antibody [E289] (ab32043)
  • Immunoprecipitation - Anti-STAT5a antibody [E289] (ab32043)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a antibody [E289] (ab32043)
  • Western blot - Anti-STAT5a antibody [E289] (ab32043)
  • Immunocytochemistry/ Immunofluorescence - Anti-STAT5a antibody [E289] (ab32043)
  • Western blot - Anti-STAT5a antibody [E289] (ab32043)
  • Western blot - Anti-STAT5a antibody [E289] (ab32043)
  • Flow Cytometry (Intracellular) - Anti-STAT5a antibody [E289] (ab32043)
  • 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

Conjugates logo Related conjugates and formulations

Alexa Fluor® 555 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free PE

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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, 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.

  • Positive control

    • WB: A431 cell lysate. IHC-P: Human squamous lung carcinoma. ICC/IF: Jurkat cells. Flow Cyt (intra): Jurkat cells.
  • 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
    For more information see here.

    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.

Properties

  • 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...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    E289
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • STATs
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-STAT5a antibody [E289] (ab194309)
    • PE Anti-STAT5a antibody [E289] (ab211686)
    • Anti-STAT5a antibody [E289] - BSA and Azide free (ab213219)
    • Alexa Fluor® 594 Anti-STAT5a antibody [E289] (ab214839)
    • Alexa Fluor® 555 Anti-STAT5a antibody [E289] (ab279325)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • A-431 whole cell lysate (ab7909)
  • Recombinant Protein

    • Recombinant Human STAT5a protein (ab84627)

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
WB (1)
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.
See IHC antigen retrieval protocols

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.

Notes
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.
See IHC antigen retrieval protocols

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

  • 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.
  • Target information above from: UniProt accession P42229 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • Unigene: 277403 Mouse
    • Unigene: 154399 Rat
    see all
  • 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

  • Western blot - Anti-STAT5a antibody [E289] (ab32043)
    Western blot - Anti-STAT5a antibody [E289] (ab32043)
    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) - Anti-STAT5a antibody [E289] (ab32043)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a antibody [E289] (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.

  • Immunocytochemistry/ Immunofluorescence - Anti-STAT5a antibody [E289] (ab32043)
    Immunocytochemistry/ Immunofluorescence - Anti-STAT5a antibody [E289] (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.

  • Flow Cytometry (Intracellular) - Anti-STAT5a antibody [E289] (ab32043)
    Flow Cytometry (Intracellular) - Anti-STAT5a antibody [E289] (ab32043)

    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). 

     

     

  • Immunoprecipitation - Anti-STAT5a antibody [E289] (ab32043)
    Immunoprecipitation - Anti-STAT5a antibody [E289] (ab32043)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a antibody [E289] (ab32043)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a antibody [E289] (ab32043)

    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).

  • Western blot - Anti-STAT5a antibody [E289] (ab32043)
    Western blot - Anti-STAT5a antibody [E289] (ab32043)
    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 - Anti-STAT5a antibody [E289] (ab32043)
    Immunocytochemistry/ Immunofluorescence - Anti-STAT5a antibody [E289] (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.

  • Western blot - Anti-STAT5a antibody [E289] (ab32043)
    Western blot - Anti-STAT5a antibody [E289] (ab32043)
    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?

  • Western blot - Anti-STAT5a antibody [E289] (ab32043)
    Western blot - Anti-STAT5a antibody [E289] (ab32043)
    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
  • Flow Cytometry (Intracellular) - Anti-STAT5a antibody [E289] (ab32043)
    Flow Cytometry (Intracellular) - Anti-STAT5a antibody [E289] (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 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. 

     

     

  • Anti-STAT5a antibody [E289] (ab32043)
    Anti-STAT5a antibody [E289] (ab32043)

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.

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (25)

Publishing research using ab32043? Please let us know so that we can cite the reference in this datasheet.

ab32043 has been referenced in 25 publications.

  • 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
  • Craig JM  et al. Prolactin Drives a Dynamic STAT5A/HDAC6/HMGN2 Cis-Regulatory Landscape Exploitable in ER+ Breast Cancer. Endocrinology 162:N/A (2021). PubMed: 33589921
  • Holloran SM  et al. Reciprocal fine-tuning of progesterone and prolactin-regulated gene expression in breast cancer cells. Mol Cell Endocrinol 511:110859 (2020). PubMed: 32407979
  • Wu G  et al. LncRNA SNHG17 aggravated prostate cancer progression through regulating its homolog SNORA71B via a positive feedback loop. Cell Death Dis 11:393 (2020). PubMed: 32447342
  • Wang C  et al. Increased SUMO-activating enzyme SAE1/UBA2 promotes glycolysis and pathogenic behavior of rheumatoid fibroblast-like synoviocytes. JCI Insight 5:N/A (2020). PubMed: 32938830
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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1-2 of 2 Abreviews or Q&A

ChIP abreview for Anti-STAT5a antibody [E289]

Inconclusive
Abreviews
Abreviews
abreview image
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)
Read More

Abcam user community

Verified customer

Submitted Nov 02 2018

Western blot abreview for Anti-STAT5a antibody [E289]

Excellent
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Mar 02 2016

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