Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-JNK2 antibody [EP1595Y] (ab76125)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1595Y] to JNK2
  • Suitable for: WB, IP, IHC-P, Flow Cyt, ELISA
  • Knockout validated
  • Reacts with: Human, Recombinant fragment

Overview

  • Product name

    Anti-JNK2 antibody [EP1595Y]
    See all JNK2 primary antibodies

  • Description

    Rabbit monoclonal [EP1595Y] to JNK2

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ELISA
    Recombinant fragment
    Flow Cyt
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human JNK2 aa 350-450 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HEK293T, MCF7, HAP1 and HeLa cell lysates. IHC-P: Human breast carcinoma tissue. IP: HeLa cell lysate. Flow Cyt: HeLa 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.

    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

Images

  • Western blot - Anti-JNK2 antibody [EP1595Y] (ab76125)
    Western blot - Anti-JNK2 antibody [EP1595Y] (ab76125)
    All lanes : Anti-JNK2 antibody [EP1595Y] (ab76125) at 1/1000 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : MAPK9 knockout HEK293T cell lysate
    Lane 3 : MCF7 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 48 kDa
    Observed band size: 48 kDa



    Lanes 1-3: Merged signal (red and green). Green - ab76125 observed at 48 kDa. Red - loading control ab8245 observed at 36 kDa. 

     ab76125 Anti-JNK2 antibody [EP1595Y]  was shown to specifically react with JNK2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266355 (knockout cell lysate ab257527) was used.  Wild-type and JNK2 knockout samples were subjected to SDS-PAGE.  ab76125 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JNK2 antibody [EP1595Y] (ab76125)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JNK2 antibody [EP1595Y] (ab76125)

    ab76125 at 1/100 dilution staining JNK2 in human breast carcinoma by Immunohistochemistry, Paraffin-embedded tissue.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Western blot - Anti-JNK2 antibody [EP1595Y] (ab76125)
    Western blot - Anti-JNK2 antibody [EP1595Y] (ab76125)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: JNK2 knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: MCF7 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab76125 observed at 54 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab76125 was shown to specifically react with JNK2 when JNK2 knockout samples were used. Wild-type and JNK2 knockout samples were subjected to SDS-PAGE. ab76125 and ab8245 (loading control to GAPDH) were diluted 1/2500 and 1/2000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 h at room temperature before imaging.

  • Immunoprecipitation - Anti-JNK2 antibody [EP1595Y] (ab76125)
    Immunoprecipitation - Anti-JNK2 antibody [EP1595Y] (ab76125)

    JNK2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit monoclonal to JNK2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab76125.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

    Band: 48kDa; JNK2

  • Western blot - Anti-JNK2 antibody [EP1595Y] (ab76125)
    Western blot - Anti-JNK2 antibody [EP1595Y] (ab76125)
    Anti-JNK2 antibody [EP1595Y] (ab76125) at 1/50000 dilution + HeLa cell lysate at 10 µg

    Secondary
    goat anti-rabbit-HRP at 1/1000 dilution

    Developed using the ECL technique.

    Predicted band size: 48 kDa
    Observed band size: 54 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 46 kDa (possible isoform)

  • Flow Cytometry - Anti-JNK2 antibody [EP1595Y] (ab76125)
    Flow Cytometry - Anti-JNK2 antibody [EP1595Y] (ab76125)
    Overlay histogram showing HeLa cells stained with ab76125 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76125, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • ELISA - Anti-JNK2 antibody [EP1595Y] (ab76125)
    ELISA - Anti-JNK2 antibody [EP1595Y] (ab76125)
    ELISA analysis of Human JNK2 recombinant protein at 250 ng/mL with ab76125. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.
  • Anti-JNK2 antibody [EP1595Y] (ab76125)
    Anti-JNK2 antibody [EP1595Y] (ab76125)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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