Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y78] to MEK2 - BSA and Azide free
  • Suitable for: ICC, WB, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-MEK2 antibody [Y78] - BSA and Azide free
    See all MEK2 primary antibodies

  • Description

    Rabbit monoclonal [Y78] to MEK2 - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    This antibody does not cross react with other MAP kinase kinase family members.

  • Tested applications

    Suitable for: ICC, WB, IHC-P, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

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

  • Positive control

    • WB: HAP1, K562, Jurkat and HEK-293T cell lysate; Mouse brain and lung lysates. ICC/IF: HeLa and wildtype HAP1 cells. IHC-P: Human prostate carcinoma tissue.

  • General notes

    ab233731 is the carrier-free version of ab32517 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    Ab233731 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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

Images

  • Western blot - Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)
    Western blot - Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)

    This data was developed using ab32517, the same antibody clone in a different buffer formulation.

    Lane 1 Wild-type HAP1 cell lysate (20 µg)

    Lane 2 MEK2 knockout HAP1 cell lysate (20 µg)

    Lane 3 Jurkat cell lysate (20 µg)

    Lane 4 K562 cell lysate (20 µg)

    Lanes 1 - 4 Merged signal (red and green). Green - ab32517 observed at 44 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab32517 was shown to specifically react with MEK2 when MEK2 knockout samples were used. Wild-type and MEK2 knockout samples were subjected to SDS-PAGE. ab32517 and ab8245 (loading control to GAPDH) were diluted 1/10 000 and 1/2000 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 1/10 000 dilution for 1 h at room temperature before imaging.

  • Immunocytochemistry - Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)
    Immunocytochemistry - Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)

    This data was developed using ab32517, the same antibody clone in a different buffer formulation.

    ab32517 staining MEK2 in wild-type HAP1 cells (top panel) and MEK2 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab32517 at 1μg/ml and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
  • Western blot - Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)
    Western blot - Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)
    All lanes : Anti-MEK2 antibody [Y78] (ab32517) at 1/10000 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : MAP2K2 knockout HEK293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 44 kDa
    Observed band size: 45 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab32517).

    Lanes 1 - 2: Merged signal (red and green). Green - ab32517 observed at 45 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab32517 was shown to react with MEK2 in wild-type HEK-293T cells in western blot with loss of signal observed in MAP2K2 knockout cell line ab266315 (MAP2K2 knockout cell lysate ab257512). Wild-type HEK-293T and MAP2K2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab32517 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)
    Western blot - Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)
    All lanes : Anti-MEK2 antibody [Y78] (ab32517) at 1/10000 dilution

    Lane 1 : Jurkat Whole Cell Lysate
    Lane 2 : Mouse Brain Tissue Lysate
    Lane 3 : Mouse Lung Tissue 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

    Performed under non-reducing conditions.

    Predicted band size: 44 kDa
    Observed band size: 44 kDa



    This data was developed using ab32517, the same antibody clone in a different buffer formulation.

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab32517 overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) at a 1:10000 dilution for 1hr at room temperature and then imaged.

  • Western blot - Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)
    Western blot - Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)
    Anti-MEK2 antibody [Y78] (ab32517) at 1/10000 dilution + Jurkat cell lysate

    Predicted band size: 44 kDa
    Observed band size: 45 kDa why is the actual band size different from the predicted?



    This data was developed using ab32517, the same antibody clone in a different buffer formulation.

  • Immunocytochemistry - Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)
    Immunocytochemistry - Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)
    This data was developed using ab32517, the same antibody clone in a different buffer formulation.Immunofluorescent staining of HeLa cells using ab32517 at a dilution of 1/250.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)
    This data was developed using ab32517, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin embedded human prostate carcinoma using ab32517 at a dilution of 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
  • Flow Cytometry - Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)
    Flow Cytometry - Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)
    This data was developed using ab32517, the same antibody clone in a different buffer formulation.Overlay histogram showing HeLa cells stained with ab32517 (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 (ab32517, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)
    Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)

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