Recombinant
RabMAb

Recombinant Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

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Overview

  • Product name

    Anti-STAT5a + STAT5b antibody [EPR16668]
    See all STAT5a+STAT5b primary antibodies

  • Description

    Rabbit monoclonal [EPR16668] to STAT5a + STAT5b

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Mouse STAT5a aa 1-250. The exact sequence is proprietary.
    Database link: P42232

  • Positive control

    • WB: Full length human Stat5a recombinant protein; recombinant protein fragment mouse stat5b; K562, Jurkat, Daudi, C6, PC-12 and NIH/3T3 whole cell lysates; mouse and rat heart lysates. IHC-P: Human tonsil, mouse liver and rat cardiac muscle tissues. ICC/IF: RAW 264.7 and C6 cells. Flow Cyt: NIH/3T3 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 ab200341 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
ICC/IF 1/500.
Flow Cyt 1/80.
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/200. Detects a band of approximately 90 kDa (predicted molecular weight: 90 kDa).

Target

Images

  • Western blot - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)
    Western blot - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)
    Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341) at 1/200 dilution + Full length human Stat5a recombinant protein at 0.01 µg

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

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


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    ab200341 show cross reactivity with STAT5a. Full length human Stat5a recombinant protein contains aa1-794 with MYC/DDDDK tag.

  • Western blot - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)
    Western blot - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)
    Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341) at 1/10000 dilution + Recombinant protein fragment mouse Stat5b at 0.01 µg

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

    Predicted band size: 90 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)
    Western blot - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)
    All lanes : Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341) at 1/1000 dilution

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

    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: 90 kDa
    Observed band size: 90 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)
    Western blot - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)
    All lanes : Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341) at 1/1000 dilution

    Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 3 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate
    Lane 4 : Mouse heart lysate
    Lane 5 : Rat heart lysate

    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: 90 kDa
    Observed band size: 90 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

    Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling STAT5b with ab200341 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Cytoplasm and nuclear staining on Human tonsil tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    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 [EPR16668] (ab200341)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

    Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling STAT5b with ab200341 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Cytoplasm and nuclear staining on mouse liver tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    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 [EPR16668] (ab200341)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

    Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling STAT5b with ab200341 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Cytoplasm and nuclear staining on rat cardiac muscle tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

  • Immunocytochemistry/ Immunofluorescence - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)
    Immunocytochemistry/ Immunofluorescence - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) cells labeling STAT5b with ab200341 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Nuclear and cytoplasm staining on RAW 264.7 cell line is observed.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab200341 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)
    Immunocytochemistry/ Immunofluorescence - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C6 (Rat glial tumor cells) cells labeling STAT5b with ab200341 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Nuclear and cytoplasm staining on C6 cell line is observed.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab200341 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • Flow Cytometry - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)
    Flow Cytometry - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

    Flow cytometric analysis of 2% paraformaldehyde-fixed NIH/3T3 (Mouse embyro fibroblast cells) cells labeling STAT5b with ab200341 at 1/80 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an 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.

References

ab200341 has not yet been referenced specifically in any publications.

Publishing research using ab200341? Please let us know so that we can cite the reference in this datasheet.

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