Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-JAK2 antibody [EPR108(2)] (ab108596)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR108(2)] to JAK2
  • Suitable for: ICC/IF, WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-JAK2 antibody [EPR108(2)]
    See all JAK2 primary antibodies

  • Description

    Rabbit monoclonal [EPR108(2)] to JAK2

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IP
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: A549, TF-1, K562, THP-1, Jurkat, NIH 3T3, Ramos, C6 and IM-9 cell lysates. ICC/IF: K562, Jurkat and Ramos 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

Images

  • Western blot - Anti-JAK2 antibody [EPR108(2)] (ab108596)
    Western blot - Anti-JAK2 antibody [EPR108(2)] (ab108596)
    All lanes : Anti-JAK2 antibody [EPR108(2)] (ab108596) at 1/1000 dilution

    Lane 1 : Wild-Type A549 cell lysate
    Lane 2 : JAK2 knockout A549 cell lysate
    Lane 3 : K562 cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 131 kDa
    Observed band size: 131 kDa



    Lanes 1- 4: Merged signal (red and green). Green - ab108596 observed at 131 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab108596 was shown to react with JAK2 in Wild-Type A549 cells in western blot. Loss of signal was observed when knockout cell line ab267113 (knockout cell lysate ab256963) was used. Wild-Type A549 and JAK2 knockout A549 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab108596 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-JAK2 antibody [EPR108(2)] (ab108596)
    Immunocytochemistry/ Immunofluorescence - Anti-JAK2 antibody [EPR108(2)] (ab108596)

    Immunocytochemistry/ Immunofluorescence analysis of K562 (Human chronic myelogenous leukemia cell line from bone marrow) cells labeling JAK2 with purified ab108596 at 1/150 dilution (8.5μg/ml). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, a goat anti-rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1/1000 dilution. Ab195889, anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used as the counter stain at 1/200 (2.5 μg/ml). PBS instead of the primary antibody was the negative control. DAPI was used as a nuclear counterstain.

  • Western blot - Anti-JAK2 antibody [EPR108(2)] (ab108596)
    Western blot - Anti-JAK2 antibody [EPR108(2)] (ab108596)
    All lanes : Anti-JAK2 antibody [EPR108(2)] (ab108596) at 1/5000 dilution (purified)

    Lane 1 : TF-1 (Human bone marrow erythroleukemia cell line) whole cell lysate
    Lane 2 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
    Lane 3 : C6 (Rat glial tumour cell line) whole cell lysate
    Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

    Predicted band size: 131 kDa
    Observed band size: 130 kDa
    why is the actual band size different from the predicted?



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

  • Western blot - Anti-JAK2 antibody [EPR108(2)] (ab108596)
    Western blot - Anti-JAK2 antibody [EPR108(2)] (ab108596)
    All lanes : Anti-JAK2 antibody [EPR108(2)] (ab108596) at 1/1000 dilution (unpurified)

    Lane 1 : TF-1 cell lysate
    Lane 2 : K562 cell lysate
    Lane 3 : THP-1 cell lysate
    Lane 4 : Jurkat cell lysate
    Lane 5 : NIH3T3 cell lysate
    Lane 6 : IM-9 cell lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 131 kDa
    Observed band size: 130 kDa why is the actual band size different from the predicted?

  • Immunocytochemistry/ Immunofluorescence - Anti-JAK2 antibody [EPR108(2)] (ab108596)
    Immunocytochemistry/ Immunofluorescence - Anti-JAK2 antibody [EPR108(2)] (ab108596)

    Immunocytochemistry/Immunofluorescence analysis of Jurkat (Human T cell leukemia cells from peripheral blood) cells labelling JAK2 with unpurified ab108596 at 1/300 (7.0 μg/mL). Cells were fixed with 4% Paraformaldehydeand permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000, 2 μg/mL) was used as the secondary antibody. Nuclei were stained with DAPI (blue).

    Confocal image showing nuclear and cytoplasmic staining on Jurkat cell line. 

  • Immunocytochemistry/ Immunofluorescence - Anti-JAK2 antibody [EPR108(2)] (ab108596)
    Immunocytochemistry/ Immunofluorescence - Anti-JAK2 antibody [EPR108(2)] (ab108596)

    Immunocytochemistry/Immunofluorescence analysis of Ramos (Human Burkitt's lymphoma) cells labelling JAK2 with unpurified 108596 at 1/300 (7.0 μg/mL). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000, 2 μg/mL) was used as the secondary antibody. Nuclei were stained with DAPI (blue).

    Confocal image showing nuclear and cytoplasmic staining on Ramos cell line.

  • Immunoprecipitation - Anti-JAK2 antibody [EPR108(2)] (ab108596)
    Immunoprecipitation - Anti-JAK2 antibody [EPR108(2)] (ab108596)

    ab108596 at 1/80 dilution (20 µg/mL) immunoprecipitating JAK2 in K562 (Human chronic myelogenous leukemia lymphoblast)  cell lysate.

    Lane 1 (input): K562(Human chronic myelogenous leukemia lymphoblast) whole cell lysate 10µg

    Lane 2 (+): K562 whole cell lysate, 350µg  + ab108596, 2µg

    Lane 3 (-): K562 cell lysate, 350µg  + rabbit IgG (ab172730), 2µg

    For western blotting, ab108596 at 1/500 dilution (3.0 µg/mL) and ab131366 VeriBlot for IP (HRP) was used at 1/1000.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Anti-JAK2 antibody [EPR108(2)] (ab108596)
    Anti-JAK2 antibody [EPR108(2)] (ab108596)

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