Recombinant
RabMAb

Anti-MEF2A + MEF2C antibody [EPR19089-34] (ab197070)

Overview

  • Product name
    Anti-MEF2A + MEF2C antibody [EPR19089-34]
    See all MEF2A+MEF2C primary antibodies
  • Description
    Rabbit monoclonal [EPR19089-34] to MEF2A + MEF2C
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human MEF2C aa 250 to the C-terminus. The exact sequence is proprietary.
    Database link: Q06413

  • Positive control
    • WB: Raji, Ramos, Daudi, C6 and RAW 264.7 whole cell lysates; Mouse cerebral cortex and brain lysates; Rat cerebral cortex, brain and spleen lysates. IHC-P: Human tonsil and colonic adenocarcinoma tissues; Mouse spleen and colon tissues; Rat spleen tissue. ICC/IF: K562 and Raji cells.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab197070 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 1/1000. Detects a band of approximately 50-60 kDa (predicted molecular weight: 51 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/250.
Flow Cyt 1/30.

Images

  • Flow Cytometry analysis of Raji(human Burkitt's lymphoma) labelling MEF2A + MEF2C with purified ab197070 at 1/30 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Alexa Fluor® 488 goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

     

  • All lanes : Anti-MEF2A + MEF2C antibody [EPR19089-34] (ab197070) at 1/1000 dilution

    Lane 1 : Recombinant Human MEF2A protein (ab152519)
    Lane 2 : Human MEF2C full length protein

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 51 kDa
    Observed band size: 55,83 kDa (why is the actual band size different from the predicted?)


    Exposure time: 1 minute


    ab197070 recognizes the full length tagged recombinant proteins MEF2A and MEF2C which have expected molecular weights of 83 and 55 kDa respectively.

    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 2% Bovine Serum Albumin before being incubated with ab197070 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • All lanes : Anti-MEF2A + MEF2C antibody [EPR19089-34] (ab197070) at 1/1000 dilution

    Lane 1 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
    Lane 2 : Ramos (Human Burkitt's lymphoma cell line) whole cell lysate
    Lane 3 : Daudi (Human Burkitt's lymphoma cell line) whole cell lysate
    Lane 4 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 51 kDa
    Observed band size: 50-60 kDa (why is the actual band size different from the predicted?)


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    MEF2C is expressed specifically in cells from B-lymphocyte lineage but not T cell lineage. Jurkat cell lysate serves as a negative control (PMID: 9798649).

    The 50-60KD bands observed are consistent with what has been described in the literature (PMID: 18450586)

  • All lanes : Anti-MEF2A + MEF2C antibody [EPR19089-34] (ab197070) at 1/5000 dilution

    Lane 1 : Mouse cerebral cortex lysate
    Lane 2 : Rat cerebral cortex lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 51 kDa
    Observed band size: 50-60 kDa (why is the actual band size different from the predicted?)


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The 50-60KD bands observed are consistent with what has been described in the literature (PMID: 18450586)

  • All lanes : Anti-MEF2A + MEF2C antibody [EPR19089-34] (ab197070) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Rat brain lysate
    Lane 3 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 51 kDa
    Observed band size: 50-60 kDa (why is the actual band size different from the predicted?)



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 1minute; Lane 2 and 3: 3minutes.

    The 50-60KD bands observed are consistent with what has been described in the literature (PMID: 18450586).

  • All lanes : Anti-MEF2A + MEF2C antibody [EPR19089-34] (ab197070) at 1/1000 dilution

    Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 2 : Raw264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 51 kDa
    Observed band size: 50-60 kDa (why is the actual band size different from the predicted?)



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 15 seconds; Lane 2: 30 seconds.

    The 50-60KD bands observed are consistent with what has been described in the literature (PMID: 18450586).

  • Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling MEF2C with ab197070 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on B lymphocytes of the Human tonsil 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.

  • Immunohistochemical analysis of paraffin-embedded Human colonic adenocarcinoma tissue labeling MEF2C with ab197070 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on lymphocytes of the colonic adenocarcinoma 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.

  • Immunohistochemical analysis of paraffin-embedded Human thymus tissue labeling MEF2C with ab197070 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on normal Human thymus is observed. Counter stained with Hematoxylin.

     

  • Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling MEF2C with ab197070 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on B lymphocytes of the mouse spleen, the T cells in the periarterial lymphatic sheath showed negative staining. 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.

  • Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling MEF2C with ab197070 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on the mouse colon epithelium, the lymphocytes on the interstitial substance showed nuclear staining. 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.

  • Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling MEF2C with ab197070 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on B lymphocytes of the rat spleen, the T cells in the periarterial lymphatic sheath showed negative staining. 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.

  • Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling MEF2C with ab197070 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on rat testis is observed. Counter stained with Hematoxylin.

     

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling MEF2C with ab197070 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear 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: ab197070 at 1/250 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.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji (Human Burkitt's lymphoma cell line) cells labeling MEF2C with ab197070 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on Raji 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: ab197070 at 1/250 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.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji (Human Burkitt's lymphoma cell line) and Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling MEF2C with ab197070 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on Raji cell line. Negative expression in Jurkat cells. 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: ab197070 at 1/250 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.

References

This product has been referenced in:
  • Harrington AJ  et al. MEF2C regulates cortical inhibitory and excitatory synapses and behaviors relevant to neurodevelopmental disorders. Elife 5:N/A (2016). WB, IHC (PFA fixed) ; Mouse . Read more (PubMed: 27779093) »

See 1 Publication for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Embryonic Heart)
Permeabilization
Yes - Tween-20
Specification
Embryonic Heart
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 06 2016

Application
Western blot
Sample
Mouse Tissue lysate - whole (Heart)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
10 µg
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 20 2016

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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