Overview

  • Product name

    Anti-MEF2C antibody [EPR19089-202] - ChIP Grade
    See all MEF2C primary antibodies
  • Description

    Rabbit monoclonal [EPR19089-202] to MEF2C - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ChIP, Flow Cyt, ICC/IF, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Positive control

    • WB: Human MEF2C recombinant protein; Raji, Jurkat, Ramos, Daudi, RAW264.7, NIH/3T3 and C6 whole cell lysates; mouse and rat brain lysate. IHC-P: Human skeletal muscle and endometrial carcinoma tissue; Mouse and rat spleen tissue. ICC/IF: Raji cells. Flow: Jurkat cells. ChIP: Chromatin prepared from HUVEC 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 ab211493 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
ChIP Use 5 µg for 25 µg of chromatin.
Flow Cyt 1/500.
ICC/IF 1/500.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Use at 1:500 (human), 1:1000 (mouse, rat) dilution.

WB 1/1000. Predicted molecular weight: 51 kDa.

Target

  • Function

    Transcription activator which binds specifically to the MEF2 element present in the regulatory regions of many muscle-specific genes. Controls cardiac morphogenesis and myogenesis, and is also involved in vascular development. Plays an essential role in hippocampal-dependent learning and memory by suppressing the number of excitatory synapses and thus regulating basal and evoked synaptic transmission. Crucial for normal neuronal development, distribution, and electrical activity in the neocortex. Necessary for proper development of megakaryocytes and platelets and for bone marrow B lymphopoiesis. Required for B-cell survival and proliferation in response to BCR stimulation, efficient IgG1 antibody responses to T-cell-dependent antigens and for normal induction of germinal center B cells. May also be involved in neurogenesis and in the development of cortical architecture (By similarity). Isoform 3 and isoform 4, which lack the repressor domain, are more active than isoform 1 and isoform 2.
  • Tissue specificity

    Expressed in brain and skeletal muscle.
  • Involvement in disease

    Defects in MEF2C are the cause of mental retardation-stereotypic movements-epilepsy and/or cerebral malformations (MRSME) [MIM:613443]. It is a disorder characterized by severe mental retardation, absent speech, hypotonia, poor eye contact and stereotypic movements. Dysmorphic features include high broad forehead with variable small chin, short nose with anteverted nares, large open mouth, upslanted palpebral fissures and prominent eyebrows. Some patients have seizures.
  • Sequence similarities

    Belongs to the MEF2 family.
    Contains 1 MADS-box domain.
    Contains 1 Mef2-type DNA-binding domain.
  • Developmental stage

    Expression is highest during the early stages of postnatal development, at later stages levels greatly decrease.
  • Domain

    The beta domain, missing in a number of isoforms, is required for enhancement of transcriptional activity.
  • Post-translational
    modifications

    Phosphorylation on Ser-59 enhances DNA binding activity (By similarity). Phosphorylation on Ser-396 is required for Lys-391 sumoylation and inhibits transcriptional activity.
    Acetylated by p300 on several sites in diffentiating myocytes. Acetylation on Lys-4 increases DNA binding and transactivation.
    Sumoylated on Lys-391 by SUMO2 but not by SUMO1 represses transcriptional activity.
    Proteolytically cleaved in cerebellar granule neurons, probably by caspase 7, following neurotoxicity. Preferentially cleaves the CDK5-mediated hyperphosphorylated form which leads to neuron apoptosis and transcriptional inactivation.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • C5DELq14.3 antibody
    • DEL5q14.3 antibody
    • MADS box transcription enhancer factor 2 polypeptide C (myocyte enhancer factor 2C) antibody
    • MADS box transcription enhancer factor 2, polypeptide C antibody
    • MEF2C antibody
    • MEF2C_HUMAN antibody
    • Myocyte enhancer factor 2C antibody
    • Myocyte specific enhancer factor 2C antibody
    • Myocyte-specific enhancer factor 2C antibody
    • OTTHUMP00000222409 antibody
    • Similar to MADS box transcription enhancer factor 2 polypeptide C antibody
    see all

Images

  • Chromatin was prepared from HUVEC cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with: 25 µg of chromatin, 5 µg of ab211493 (red), and 20 µl of protein A/G sepharose beads slurry (10 µl of sepharose A beads + 10 µl of sepharose G beads). 5 μg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (SYBR green chemistry).

    ChIP was performed according to the literature (PMID: 26923194).

  • All lanes : Anti-MEF2C antibody [EPR19089-202] - ChIP Grade (ab211493) at 1/1000 dilution

    Lane 1 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
    Lane 2 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
    Lane 3 : C6 (rat glial tumor glial cell), whole cell lysate
    Lane 4 : Mouse brain lysate
    Lane 5 : Rat brain lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lanes 1-3 & 5 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
    Lane 4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

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



    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    Exposure time:

    Lanes 1-2,4: 5 minutes

    Lane 3: 15 seconds

    Lane 5: 26 seconds

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilsed Jurkat (Human T cell leukemia T lymphocyte, Left) / Raji (Human Burkitt's lymphoma B lymphocyte, Right) cell lines labelling MEF2C with ab211493 at 1/500 dilution (Red) compared with the isotype control Rabbit monoclonal IgG (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG Alexa Fluor® 488 (ab150077), at 1/2000 dilution was used as the secondary antibody.

    Negative control: Jurkat (PMID: 27876533).

  • All lanes : Anti-MEF2C antibody [EPR19089-202] - ChIP Grade (ab211493) at 1/1000 dilution

    Lane 1 : Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate 20 µg
    Lane 2 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate 20 µg
    Lane 3 : Ramos (human Burkitt's lymphoma B lymphocyte), whole cell lysate 20µg
    Lane 4 : Daudi (human Burkitt's lymphoma lymphoblast), whole cell lysate 20 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

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



    Blocking and diluting buffer and concentration: 5% NFDM/TBST 

    Exposure time: 92 seconds.

    The expression profile observed is consistent with the literature (PMID: 18450586). Negative control: Jurkat (PMID: 27876533)

  • All lanes : Anti-MEF2C antibody [EPR19089-202] - ChIP Grade (ab211493) at 1/1000 dilution

    Lane 1 : Human MEF2C recombinant protein (aa271-493) 10 ng
    Lane 2 : Human MEF2A recombinant protein (aa1-499, ab204772) 10 ng

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

    Predicted band size: 51 kDa
    Observed band size: 26 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST
    Exposure time: 3 seconds. 

  • Immunohistochemical analysis of paraffin-embedded rat spleen tissue labelling MEF2C with ab211493 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining in B lymphocytes of rat spleen but not in T cells in the periarterial lymphatic sheath is observed (PMID: 8506376, PMID 15703219). Counterstained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP)ready to use.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labelling MEF2C with ab211493 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining in B lymphocytes of mouse spleen but not in T cells in the periarterial lymphatic sheath is observed (PMID: 8506376, PMID 15703219). Counterstained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP)ready to use.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded human endometrial carcinoma tissue labelling MEF2C with ab211493 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining in leukocytes but not in tumor cells of human endometrial carcinoma is observed (PMID: 8506376, PMID 15703219). Counterstained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP)ready to use.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labelling MEF2C with ab211493 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining in human skeletal muscle cells is observed(PMID: 8506376, PMID 15703219). Counterstained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP)ready to use.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilised Raji (human Burkitt's lymphoma B lymphocyte) cells labelling MEF2C with ab211493 at 1/500 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution (green). Confocal image showing nuclear staining in Raji cell line. Negative control: Jurkat (PMID: 27876533). DAPI was used as the nuclear counterstain, and the Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889 antibody was used as a counterstain at 1/200 dilution.

    The negetive controls are as follows:
    -ve control 1: ab197070 on jurkat (human T cell leukemia cell line from peripheral blood) cells.
    -ve control 2: Jurkat cells stained with DAPI.
    -ve control 3: Merged negetive contol images.

References

ab211493 has not yet been referenced specifically in any publications.

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