RecombinantRabMAb

Recombinant Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR16671-40] to STAT5a + STAT5b
  • Suitable for: IHC-P, WB, ICC/IF, Flow Cyt, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-STAT5a + STAT5b antibody [EPR16671-40]

  • Description

    Rabbit monoclonal [EPR16671-40] to STAT5a + STAT5b

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WB, ICC/IF, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Mouse STAT5a aa 600 to the C-terminus. The exact sequence is proprietary. Also SwissProt ID: P42232 (mouse)
    Database link: P42230

  • Positive control

    • WB: Human STAT5A full length recombinant protein; K562, Jurkat and Daudi cell lysates; Human fetal heart and kidney lysates; mouse heart, kidney, spleen lysates; rat brain, heart, spleen lysates; RAW 264.7, PC12, NIH/3T3 whole cell lysates. IHC-P: Human tonsils, mouse spleen and rat spleen tissue. IF, Flow, IP: K562 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab194898 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
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/2000. Detects a band of approximately 91 kDa (predicted molecular weight: 91 kDa).
ICC/IF 1/250.
Flow Cyt 1/80.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 
IP 1/100.

Target

Images

  • Western blot - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
    Western blot - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
    All lanes : Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898) at 1/20000 dilution

    Lane 1 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysates
    Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates
    Lane 3 : Daudi (Human Burkitt's lymphoma cell line) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 91 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
    Immunocytochemistry/ Immunofluorescence - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling STAT5a + STAT5b with ab194898 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing both cytoplasmic and nuclear staining on K562 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor® 594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:
    1. ab194898 at 1/250 dilution followed by ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

  • Western blot - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
    Western blot - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
    All lanes : Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898) at 1/2000 dilution

    Lane 1 : Human fetal heart lysates
    Lane 2 : Human fetal kidney lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 91 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
    Western blot - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
    Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898) at 1/1000 dilution + Human STAT5A full length recombinant protein at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 91 kDa



    Recombinant human C-terminal Myc/DDK tagged full length STAT5A from a commercial source.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
    Western blot - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
    All lanes : Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898) at 1/2000 dilution

    Lane 1 : mouse heart lysates
    Lane 2 : mouse kidney lysates
    Lane 3 : mouse spleen lysates
    Lane 4 : rat brain lysates
    Lane 5 : rat heart lysates
    Lane 6 : rat spleen lysates
    Lane 7 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates
    Lane 8 : PC12 (Rat adrenal gland pheochromocytoma) whole cell lysates
    Lane 9 : NIH3T3 (Mouse embyro fibroblast cells) whole cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 91 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling STAT5a + STAT5b using ab194898 at 1/2000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 was used as secondary. Counter stained with Hematoxylin.
    Inset image: negative control obtained using PBS instead of ab194898.
    Nuclear and weak cytoplasmic staining on lymphocytes of human tonsil is observed.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

    Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling STAT5a + STAT5b using ab194898 at 1/2000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 was used as secondary. Counter stained with Hematoxylin.
    Inset image: negative control obtained using PBS instead of ab194898.
    Nuclear and weak cytoplasmic staining on lymphocytes of mouse spleen is observed.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

    Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling STAT5a + STAT5b using ab194898 at 1/2000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 was used as secondary. Counter stained with Hematoxylin.
    Inset image: negative control obtained using PBS instead of ab194898.
    Nuclear and weak cytoplasmic staining on lymphocytes of rat spleen is observed.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
    Flow Cytometry - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

    Flow cytometric analysis of 2% paraformaldehyde-fixed K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling STAT5a + STAT5b with ab194898 at 1/80 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

  • Immunoprecipitation - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
    Immunoprecipitation - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

    STAT5a and STAT5b were immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) cells with ab194898 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab194898 at 1/1000 dilution. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated, was used as secondary antibody at 1/1000 dilution.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  • Immunoprecipitation - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
    Immunoprecipitation - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

    STAT5a + STAT5b were co-immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) cells with ab194898 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab32043 (Rabbit monoclonal [E289] to STAT5a) at 1/1000 dilution. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated, was used as secondary antibody at 1/1000 dilution.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  • Immunoprecipitation - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
    Immunoprecipitation - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

    STAT5a + STAT5b were co-immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) cells with ab194898 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab178941 (Rabbit monoclonal [EPR16671] to STAT5b) at 1/1000 dilution. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated, was used as secondary antibody at 1/1000 dilution.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  • Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
    Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

References (6)

Publishing research using ab194898? Please let us know so that we can cite the reference in this datasheet.

ab194898 has been referenced in 6 publications.

  • Høyer KL  et al. The acute effects of growth hormone in adipose tissue is associated with suppression of antilipolytic signals. Physiol Rep 8:e14373 (2020). PubMed: 32073221
  • Zhang Y Effect of iron chelation therapy on EPO-STAT5 signalling pathway and EPO resistance in iron-overloaded low-risk myelodysplastic syndrome patients. Hematology 25:1-10 (2020). PubMed: 31838956
  • Aikawa T  et al. Quizartinib, a selective FLT3 inhibitor, maintains antileukemic activity in preclinical models of RAS-mediated midostaurin-resistant acute myeloid leukemia cells. Oncotarget 11:943-955 (2020). PubMed: 32215183
  • Ding L  et al. Deletion of Cdkn1b in ACI rats leads to increased proliferation and pregnancy-associated changes in the mammary gland due to perturbed systemic endocrine environment. PLoS Genet 15:e1008002 (2019). PubMed: 30893315
  • You YH  et al. MicroRNA-153 promotes brain-derived neurotrophic factor and hippocampal neuron proliferation to alleviate autism symptoms through inhibition of JAK-STAT pathway by LEPR. Biosci Rep 39:N/A (2019). PubMed: 30975733
  • Mitchell J  et al. Validation of a Preclinical Model of Diethylnitrosamine-Induced Hepatic Neoplasia in Yucatan Miniature Pigs. Oncology 91:90-100 (2016). PubMed: 27305144

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-STAT5a + STAT5b antibody [EPR16671-40]

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (spleen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
Permeabilization
No
Specification
spleen
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 Jul 31 2019

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-STAT5a + STAT5b antibody [EPR16671-40]

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (tonsil)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
Permeabilization
No
Specification
tonsil
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 Jul 31 2019

Western blot abreview for Anti-STAT5a + STAT5b antibody [EPR16671-40]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Tissue lysate - whole (human sceletal muscle)
Gel Running Conditions
Reduced Denaturing (WES simple western)
Loading amount
1 µg
Specification
human sceletal muscle
Blocking step
antibody dilution from protein simple as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 23°C
Read More

Helle Zibrandtsen

Verified customer

Submitted Mar 07 2018

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