Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)

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Western blot - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
  • Western blot - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
  • Immunocytochemistry - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
  • Western blot - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
  • Western blot - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
  • Western blot - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
  • Immunocytochemistry - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
  • Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6528(N)] to MAPK 12 - BSA and Azide free
  • Suitable for: WB, ICC
  • Knockout validated
  • Reacts with: Rat, Human

Overview

  • Product name

    Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free
    See all MAPK 12 primary antibodies

  • Description

    Rabbit monoclonal [EPR6528(N)] to MAPK 12 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICCmore details

  • Species reactivity

    Reacts with: Rat, Human

  • Immunogen

    Synthetic peptide within Human MAPK 12 aa 50-150. The exact sequence is proprietary.
    Database link: P53778

  • Positive control

    • WB: Human skeletal muscle lysate; HEK-293T, HeLa, K562, C6 and PC-12 whole cell lysates; Rat heart, spleen and muscle lysates. ICC/IF: A673 and K562 cells.

  • General notes

    Ab251440 is the carrier-free version of ab205926. 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 <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab251440 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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

Applications

Our Abpromise guarantee covers the use of ab251440 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
WB Use at an assay dependent concentration. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).
ICC Use at an assay dependent concentration.

Target

  • Function

    Responds to activation by environmental stress and pro-inflammatory cytokines by phosphorylating downstream targets. Plays a role in myoblast differentiation and also in the down-regulation of cyclin D1 in response to hypoxia in adrenal cells suggesting MAPK12 may inhibit cell proliferation while promoting differentiation.

  • Tissue specificity

    Highly expressed in skeletal muscle and heart.

  • Sequence similarities

    Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
    Contains 1 protein kinase domain.

  • Domain

    The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.

  • Post-translational
    modifications

    Dually phosphorylated on Thr-183 and Tyr-185, which activates the enzyme.

  • Cellular localization

    Cytoplasm. Mitochondrion. Mitochondrial when associated with SH3BP5.
  • Information by UniProt

  • Database links

    • Alternative names

      • ERK 6 antibody
      • ERK-6 antibody
      • ERK6 antibody
      • Extracellular signal-regulated kinase 6 antibody
      • MAP kinase 12 antibody
      • MAP kinase p38 gamma antibody
      • MAPK 12 antibody
      • Mapk12 antibody
      • Mitogen Activated Protein Kinase 12 antibody
      • Mitogen activated protein kinase p38 gamma antibody
      • Mitogen-activated protein kinase 12 antibody
      • Mitogen-activated protein kinase p38 gamma antibody
      • MK12_HUMAN antibody
      • P38 GAMMA antibody
      • P38GAMMA antibody
      • PRKM12 antibody
      • SAPK 3 antibody
      • SAPK-3 antibody
      • SAPK3 antibody
      • Stress Activated Protein Kinase 3 antibody
      • Stress-activated protein kinase 3 antibody
      see all

    Images

    • Western blot - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
      Western blot - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)

      This data was developed using ab205926, the same antibody clone in a different buffer formulation.

      Lane 1: Wild type HAP1 whole cell lysate (0 µg)

      Lane 2: empty knockout HAP1 whole cell lysate (20 µg)

      Lane 3: MAPK 12 whole cell lysate (0 µg)

      Lanes 1 - 3: Merged signal (red and green). Green - ab205926 observed at 45 kDa. Red - loading control, ab18058, observed at 124 kDa.

    • Western blot - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
      Western blot - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
      All lanes : Anti-MAPK 12 antibody [EPR6528(N)] (ab205926) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
      Lane 2 : MAPK12 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
      Lane 3 : Human skeletal muscle 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

      Predicted band size: 42 kDa
      Observed band size: 42 kDa



      This data was developed using ab205926, the same antibody clone in a different buffer formulation.

      Lanes 1-3: Merged signal (red and green). Green - ab205926 observed at 42 kDa. Red - loading control ab8245 observed at 36 kDa.

       ab205926 Anti-MAPK 12 antibody [EPR6528(N)] was shown to specifically react with MAPK 12 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266280 (knockout cell lysate ab258041) was used. Wild-type and MAPK 12 knockout samples were subjected to SDS-PAGE. ab205926 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.

    • Immunocytochemistry - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
      Immunocytochemistry - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
      This data was developed using ab205926, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A673 (Human muscle Ewing's Sarcoma cell line) cells labeling MAPK 12 with ab205926 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and cytoplasmic staining on A673 cell line. The nuclear counterstain 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 1/1000 dilution (red). The negative controls are as follows:- -ve control 1: ab205926 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 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 1/1000 dilution.
    • Western blot - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
      Western blot - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
      All lanes : Anti-MAPK 12 antibody [EPR6528(N)] (ab205926) at 1/10000 dilution

      Lane 1 : Human skeletal muscle tissue lysate
      Lane 2 : Human fetal kidney tissue lysate
      Lane 3 : Human fetal liver tissue lysate
      Lane 4 : Human fetal skin tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

      Predicted band size: 42 kDa
      Observed band size: 42 kDa


      Exposure time: 3 minutes


      This data was developed using ab205926, the same antibody clone in a different buffer formulation.

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
      Western blot - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
      All lanes : Anti-MAPK 12 antibody [EPR6528(N)] (ab205926) at 1/1000 dilution

      Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
      Lane 2 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

      Predicted band size: 42 kDa
      Observed band size: 42 kDa


      Exposure time: 3 minutes


      This data was developed using ab205926, the same antibody clone in a different buffer formulation.

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
      Western blot - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
      All lanes : Anti-MAPK 12 antibody [EPR6528(N)] (ab205926) at 1/1000 dilution

      Lane 1 : Rat heart tissue lysate
      Lane 2 : Rat spleen tissue lysate
      Lane 3 : C6 (Rat glial tumor cell line) whole cell lysate
      Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
      Lane 5 : Rat muscle tissue lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

      Predicted band size: 42 kDa
      Observed band size: 42 kDa



      This data was developed using ab205926, the same antibody clone in a different buffer formulation.

      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: Lanes 1-2: 3 minutes; Lane 3-4: 1 minute; Lane 5: 30 seconds.

    • Immunocytochemistry - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
      Immunocytochemistry - Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
      This data was developed using ab205926, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling MAPK 12 with ab205926 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and cytoplasmic staining on K562 cell line. The nuclear counterstain 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 1/1000 dilution (red). The negative controls are as follows:- -ve control 1: ab205926 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 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 1/1000 dilution.
    • Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)
      Anti-MAPK 12 antibody [EPR6528(N)] - BSA and Azide free (ab251440)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab251440? Please let us know so that we can cite the reference in this datasheet.

    ab251440 has not yet been referenced specifically in any publications.

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