Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR17340] to MEK3 + MEK6 - BSA and Azide free
  • Suitable for: Flow Cyt, WB, IP, IHC-P, ICC
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free
    See all MEK3 + MEK6 primary antibodies

  • Description

    Rabbit monoclonal [EPR17340] to MEK3 + MEK6 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

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

  • Immunogen

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

  • General notes

    Ab251322 is the carrier-free version of ab200831. 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

    ab251322 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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)
    Western blot - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)
    All lanes : Anti-MEK3 + MEK6 antibody [EPR17340] (ab200831) at 1/1000 dilution

    Lane 1 : Mouse spleen lysate at 10 µg
    Lane 2 : Rat spleen lysate at 10 µg
    Lane 3 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 4 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 6 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

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

    Predicted band size: 39, 37 kDa
    Observed band size: 34,37,39 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    This data was developed using ab200831, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)

    This data was developed using ab200831, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling MEK3 + MEK6 with ab200831 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and cytoplasmic 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.
  • Western blot - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)
    Western blot - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)

    This data was developed using ab200831, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Recombinant fragment of Human MEK6 protein contains aa139-334 with His-Tag®(22kDa).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)

    This data was developed using ab200831, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling MEK3 + MEK6 with ab200831 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and cytoplasmic staining on Human cervix carcinoma 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.
  • Western blot - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)
    Western blot - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)
    All lanes : Anti-MEK3 + MEK6 antibody [EPR17340] (ab200831) at 1/1000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate
    Lane 3 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
    Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 39, 37 kDa
    Observed band size: 34,37,39 kDa why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    This data was developed using ab200831, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)

    This data was developed using ab200831, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling MEK3 + MEK6 with ab200831 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and cytoplasmic staining on Human spleen 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.
  • Western blot - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)
    Western blot - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)
    Anti-MEK3 + MEK6 antibody [EPR17340] (ab200831) at 1/10000 dilution + Human fetal liver tissue lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 39, 37 kDa
    Observed band size: 34,37,39 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    This data was developed using ab200831, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)
    Immunocytochemistry - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)

    This data was developed using ab200831, the same antibody clone in a different buffer formulation.

     

    Immunocytochemistry analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (human hepatocellular carcinoma) cells labeling MEK3 + MEK6 with ab200831 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Nuclear and cytoplasmic staining on HepG2 cell line is observed. 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 1/500 dilution (red).
    The negative controls are as follows:-
    -ve control 1: ab200831 at 1/1000 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-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)
    Flow Cytometry - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)

    This data was developed using ab200831, the same antibody clone in a different buffer formulation.

    Flow cytometry analysis of HeLa cells labelling MEK3 + MEK6 (red) with purified ab200831 at dilution of 1/150. The secondary antibody used was Alexa Fluor® 488 goat-anti-rabbit IgG (1/2000). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

  • Immunoprecipitation - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)
    Immunoprecipitation - Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)
    This data was developed using ab200831, the same antibody clone in a different buffer formulation.MEK3 + MEK6 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab200831 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab200831 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.Lane 1: HeLa whole cell lysate 10ug (Input). Lane 2: ab200831 IP in HeLa whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200831 in Jurkat whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.Exposure time: 30 seconds.
  • Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)
    Anti-MEK3 + MEK6 antibody [EPR17340] - BSA and Azide free (ab251322)

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