Recombinant
RabMAb

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

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

    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 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

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

References

This product has been referenced in:
  • Mitchell J  et al. Validation of a Preclinical Model of Diethylnitrosamine-Induced Hepatic Neoplasia in Yucatan Miniature Pigs. Oncology 91:90-100 (2016). Read more (PubMed: 27305144) »
See 1 Publication for this product

Customer reviews and Q&As

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

Helle Zibrandtsen

Verified customer

Submitted Mar 07 2018

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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