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    stat5-phospho-y694-antibody-e208-ab32364.pdf

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RecombinantRabMAb

Recombinant Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)

  • Datasheet
  • SDS
Reviews (10)Q&A (2)References (50)

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Western blot - Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)
  • ELISA - Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)
  • Western blot - Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)
  • Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E208] to STAT5 (phospho Y694)
  • Suitable for: WB, IHC-P, ELISA
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-STAT5 (phospho Y694) antibody [E208]
    See all STAT5 primary antibodies
  • Description

    Rabbit monoclonal [E208] to STAT5 (phospho Y694)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ELISAmore details
    Unsuitable for: ICC/IF or IP
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A431 cell lysate treated with epidermal growth factor. IHC-P: Human breast carcinoma tissue.
  • 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.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

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

    E208
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • STATs
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • STATs
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • STAT family
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Alternative Versions

    • Anti-STAT5 (phospho Y694) antibody [E208] - BSA and Azide free (ab219607)
  • Compatible Secondaries

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

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

    • A-431 nuclear extract lysate (EGF treated) (ab14635)
    • A-431 whole cell lysate (ab7909)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32364 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 (4)
1/1000. Detects a band of approximately 90 kDa (predicted molecular weight: 90 kDa).
IHC-P (5)
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ELISA
Use at an assay dependent concentration.
Notes
WB
1/1000. Detects a band of approximately 90 kDa (predicted molecular weight: 90 kDa).
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ELISA
Use at an assay dependent concentration.
Application notes
Is unsuitable for ICC/IF or IP.

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: 6777 Human
    • Entrez Gene: 20850 Mouse
    • Entrez Gene: 20851 Mouse
    • Omim: 601511 Human
    • Omim: 604260 Human
    • SwissProt: P42229 Human
    • SwissProt: P51692 Human
    • SwissProt: P42230 Mouse
    • SwissProt: P42232 Mouse
    • Unigene: 437058 Human
    • Unigene: 277403 Mouse
    see all
  • Alternative names

    • MGF antibody
    • Signal transducer and activator of transcription 5A antibody
    • Signal Transducer and Activator of Transcription 5B antibody
    • STA5A_HUMAN antibody
    • STAT 5A antibody
    • STAT 5B antibody
    • STAT5 antibody
    • STAT5A antibody
    • STAT5B antibody
    • Transcription factor STAT5A antibody
    • Transcription factor STAT5B antibody
    see all

Images

  • Western blot - Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)
    Western blot - Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)
    All lanes : Anti-STAT5 (phospho Y694) antibody [E208] (ab32364) at 1/1000 dilution (purified)

    Lane 1 : A431 cell lysate, untreated
    Lane 2 : A431 cell lysate, treated with Epidermal growth factor

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 90 kDa
    Observed band size: 90 kDa



    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human infiltrating duct carcinoma of breast tissue labelling STAT5 (phospho Y694) with purified ab32364 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • ELISA - Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)
    ELISA - Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)

    Serially diluted ab32364 was bound to immobilised phospho peptide (P) - or control peptides ((C), 1 microgram x mL-1). The antibody was detected by HRP-labelled goat anti-rabbit IgG (ab97080; diluted 1: 50,000) and signal was developed with TMB substrate

  • Western blot - Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)
    Western blot - Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)
    All lanes : Anti-STAT5 (phospho Y694) antibody [E208] (ab32364) at 1/1000 dilution (unpurified)

    Lane 1 : A431 cell lysate, untreated
    Lane 2 : A431 cell lysate, treated with EGF
    Lane 3 : A431 cell lysate, treated with phosphatase

    Predicted band size: 90 kDa
    Observed band size: 90 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma using unpurified ab32364 at a 1/50 dilution.

  • Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)
    Anti-STAT5 (phospho Y694) antibody [E208] (ab32364)

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

  • SDS download

  • Datasheet download

    Download

References (50)

Publishing research using ab32364? Please let us know so that we can cite the reference in this datasheet.

ab32364 has been referenced in 50 publications.

  • Gao H  et al. Characterization of murine mammary stem/progenitor cells in a D-galactose-induced aging model. Aging (Albany NY) 13:11762-11773 (2021). PubMed: 33878032
  • Li R  et al. Identification and In Vitro Functional Verification of Two Novel Mutations of GHR Gene in the Chinese Children with Laron Syndrome. Front Endocrinol (Lausanne) 12:605736 (2021). PubMed: 33912130
  • Mäkinen A  et al. IGF2BP3 Associates with Proliferative Phenotype and Prognostic Features in B-Cell Acute Lymphoblastic Leukemia. Cancers (Basel) 13:N/A (2021). PubMed: 33805930
  • Wang S  et al. Long non-coding RNA SNHG1 suppresses cell migration and invasion and upregulates SOCS2 in human gastric carcinoma. Biochem Biophys Rep 27:101052 (2021). PubMed: 34179518
  • Dong Z  et al. CCL26 regulates the proportion of CD4+CD25+FOXP3+ Tregs and the production of inflammatory factors in peripheral blood mononuclear cells following acute ischemic stroke via the STAT5 pathway. Exp Ther Med 20:3343-3351 (2020). PubMed: 32855707
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-10 of 12 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-STAT5 (phospho Y694) antibody [E208]

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (breast carcinoma)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
Permeabilization
No
Specification
breast carcinoma
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: RT°C
Fixative
10% NBF
Read More

Abcam user community

Verified customer

Submitted Mar 14 2019

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-STAT5 (phospho Y694) antibody [E208]

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (lymph node)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
Permeabilization
No
Specification
lymph node
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: RT°C
Fixative
10% NBF
Read More

Abcam user community

Verified customer

Submitted Mar 14 2019

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-STAT5 (phospho Y694) antibody [E208]

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Cynomolgus monkey Tissue sections (Colon)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA
Permeabilization
No
Specification
Colon
Blocking step
Protein Block (ab64226) as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Dec 28 2017

Western blot abreview for Anti-STAT5 (phospho Y694) antibody [E208]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (mammary epithelial cells)
Gel Running Conditions
Reduced Denaturing (8%)
Loading amount
40 µg
Specification
mammary epithelial cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Flavia Ghiraldini

Verified customer

Submitted Jul 05 2017

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-STAT5 (phospho Y694) antibody [E208]

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Mesenteric LN)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA pH 9.0
Permeabilization
No
Specification
Mesenteric LN
Blocking step
Casein in PBS as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Nov 30 2016

Immunocytochemistry/ Immunofluorescence abreview for Anti-STAT5 (phospho Y694) antibody [E208]

Good
Abreviews
Abreviews
This product is known to not work in this application or species.
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
Sample
Human Cell (Hela cells)
Specification
Hela cells
Permeabilization
Yes - citrate buffer, pH 6.0
Fixative
Methanol
Read More

Abcam user community

Verified customer

Submitted Mar 23 2015

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-STAT5 (phospho Y694) antibody [E208]

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer pH 6,0
Sample
Human Tissue sections (skin)
Specification
skin
Permeabilization
Yes - 0,1% Saponin/PBS
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Mar 17 2015

Western blot abreview for Anti-STAT5 (phospho Y694) antibody [E208]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
15 µg
Gel Running Conditions
Reduced Non-Denaturing (Native) (5-16% gel)
Sample
Human Cell lysate - whole cell (Jurkat and Hela cells)
Specification
Jurkat and Hela cells
Treatment
see below
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Mar 17 2015

Question

Inquiry: Which antibody do you suggest to detect mouse pSTAT5B (Y699)? I could only find pSTAT5 antibodies which detect phosphorylation at Y694, which more specific to STAT5A in your catalog.

Read More

Abcam community

Verified customer

Asked on Aug 02 2016

Answer

I can confirm that the sequence around the phosphorylation site for STAT5A phospho Y694 and STAT5B phospho Y699 is 100% homolog:


5A: VDGYPVKP

5B: VDGYPVKP


Therefore any antibody that binds STAT5 (phospho Y694) will also bind to STAT5 (phospho Y699). This means that any antibody that is named STAT5 (phospho Y694) in our catalog is suitable for your purposes.


Please note that it is impossible to generate an antibody that only binds STAT5B (phospho Y699) due to the homology with STAT5A (phospho Y694).


Read More

Abcam Scientific Support

Answered on Aug 02 2016

Question

To whom this may concern, I am working with a b32364 and having problems getting any detectable protein using western blotting from either human primary macrophages or murine bone derived macrophages.  I would like to use a positive control for the blot and was hoping that you would be able to recommend a product for both murine and human detection.    

Read More

Abcam community

Verified customer

Asked on Nov 22 2011

Answer

Thank you for contacting us. For a positive control for human cells, I recommend the nuclear lysate of EGF-treated A431 cells which we have in our catalogue as ab14635: Click here (or use the following: https://www.abcam.com/A431-nuclear-extract-lysate-EGF-treated-ab14635.html). For mouse cells, I recommend a lysate of 3T3 cells treated with leukemia inhibitory facotr (LIF), as seen in the western blot shown on the datasheet for ab13539, but we do not have this lysate as a product. There may be other treatments of mouse cells that will induce the phosphorylation, but we are not aware of them. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Abcam Scientific Support

Answered on Nov 22 2011

1-10 of 12 Abreviews or Q&A

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