Product nameAnti-MEF2A (phospho T312) antibody
See all MEF2A primary antibodies
DescriptionRabbit polyclonal to MEF2A (phospho T312)
Tested applicationsSuitable for: IHC-P, ELISA, WB, IP, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
Synthetic phosphopeptide derived from human MEF2A around the phosphorylation site of Threonine 312.
- Breast carcinoma tissue; NIH/3T3 cell lysate treated with PMA. IF: Formaldehyde fixed HeLa cell line
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride
Without Mg2+ and Ca2+
Concentration information loading...
PurityImmunogen affinity purified
Purification notesThe antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
Our Abpromise guarantee covers the use of ab30644 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/50 - 1/100.|
|WB||1/500 - 1/1000. Detects a band of approximately 65 kDa (predicted molecular weight: 55 kDa).|
|IP||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 1 µg/ml.|
FunctionTranscriptional activator which binds specifically to the MEF2 element, 5'-YTA[AT](4)TAR-3', found in numerous muscle-specific genes. Also involved in the activation of numerous growth factor- and stress-induced genes. Mediates cellular functions not only in skeletal and cardiac muscle development, but also in neuronal differentiation and survival. Plays diverse roles in the control of cell growth, survival and apoptosis via p38 MAPK signaling in muscle-specific and/or growth factor-related transcription. In cerebellar granule neurons, phosphorylated and sumoylated MEF2A represses transcription of NUR77 promoting synaptic differentiation.
Tissue specificityIsoform MEF2 and isoform MEFA are expressed only in skeletal and cardiac muscle and in the brain. Isoform RSRFC4 and isoform RSRFC9 are expressed in all tissues examined.
Involvement in diseaseDefects in MEF2A might be a cause of autosomal dominant coronary artery disease 1 with myocardial infarction (ADCAD1) [MIM:608320].
Sequence similaritiesBelongs to the MEF2 family.
Contains 1 MADS-box domain.
Contains 1 Mef2-type DNA-binding domain.
modificationsConstitutive phosphorylation on Ser-408 promotes Lys-403 sumoylation thus preventing acetylation at this site. Dephosphorylation on Ser-408 by PPP3CA upon neuron depolarization promotes a switch from sumoylation to acetylation on residue Lys-403 leading to inhibition of dendrite claw differentiation. Phosphorylation on Thr-312 and Thr-319 are the main sites involved in p38 MAPK signaling and activate transcription. Phosphorylated on these sites by MAPK14/p38alpha and MAPK11/p38beta, but not by MAPK13/p38delta nor by MAPK12/p38gamma. Phosphorylation on Ser-408 by CDK5 induced by neurotoxicity inhibits MEF2A transcriptional activation leading to apoptosis of cortical neurons. Phosphorylation on Thr-312, Thr-319 and Ser-355 can be induced by EGF.
Sumoylation on Lys-403 is enhanced by PIAS1 and represses transcriptional activity. Phosphorylation on Ser-408 is required for sumoylation. Has no effect on nuclear location nor on DNA binding. Sumoylated by SUMO1 and, to a lesser extent by SUMO2 and SUMO3. PIASx facilitates sumoylation in postsynaptic dendrites in the cerebellar cortex and promotes their morphogenesis.
Acetylation on Lys-403 activates transcriptional activity. Acetylated by p300 on several sites in diffentiating myocytes. Acetylation on Lys-4 increases DNA binding and transactivation (By similarity). Hyperacetylation by p300 leads to enhanced cardiac myocyte growth and heart failure.
Proteolytically cleaved in cerebellar granule neurons on several sites by caspase 3 and caspase 7 following neurotoxicity. Preferentially cleaves the CDK5-mediated hyperphosphorylated form which leads to neuron apoptosis and transcriptional inactivation.
- Information by UniProt
- ADCAD1 antibody
- MADS box transcription enhancer factor 2, polypeptide A (myocyte enhancer factor 2A) antibody
- MEF2 antibody
Anti-MEF2A (phospho T312) antibody (ab30644) at 1/1000 dilution + Mouse liver tissue lysate - whole at 40 µg
HRP-conjugated goat anti-rabbit IgG polyclonal at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 55 kDa
Exposure time: 1 minute
Immunohistochemical analysis of paraffin-embedded breast carcinoma. Left: using ab30644 (10ug/ml); right: the same antibody preincubated with synthesized phosphopeptide.
All lanes : Anti-MEF2A (phospho T312) antibody (ab30644) at 1 µg/ml
Lane 1 : Untreated NIH/3T3 cells (5-30ug).
Lane 2 : NIH/3T3 cells treated with PMA (5-30ug)
Predicted band size: 55 kDa
Observed band size: 65 kDa why is the actual band size different from the predicted?
ICC/IF image of ab30644 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab30644, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
This product has been referenced in:
- Ohashi A et al. CAPE increases the expression of SOD3 through epigenetics in human retinal endothelial cells. J Clin Biochem Nutr 61:6-13 (2017). Read more (PubMed: 28751803) »
- Willemen HL et al. A novel p38 MAPK docking-groove-targeted compound is a potent inhibitor of inflammatory hyperalgesia. Biochem J 459:427-39 (2014). Read more (PubMed: 24517375) »