Key features and details
- Assay type: Semi-quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 3 hr 30 min
- Sample type: Nuclear Extracts
- Sensitivity: 600 ng/well
Product nameMEF2 Transcription Factor Assay Kit (colorimetric)
Sample typeNuclear Extracts
Assay time3h 30m
Species reactivityReacts with: Human
MEF2 Transcription Factor Assay Kit (Colorimetric) (ab207213) is a high throughput assay to quantify MEF2 activation in nuclear extracts. This assay combines a quick ELISA format with a sensitive and specific non-radioactive assay for transcription factor activation.
A specific double stranded DNA sequence containing the MEF2 consensus binding site has been immobilized onto a 96-well plate. Active MEF2 present in the nuclear extract specifically binds to the oligonucleotide. MEF2 is detected by a primary antibody that recognizes an epitope of MEF2 accessible only when the protein is activated and bound to its target DNA. An HRP-conjugated secondary antibody provides sensitive colorimetric readout at OD 450 nm. This product detects only human MEF2.
Key performance and benefits:
- Assay time: 3.5 hours (cell extracts preparation not included).
- Detection limit: < 0.6 µg nuclear extract/well.
- Detection range: 0.6 – 10 µg nuclear extract/well.
The transmission of extracellular signals into intracellular responses is a complex process that often involves the activity of mitogen-activated protein kinases (MAPKs). The MAPK pathway is a three kinase cascade consisting of a MAPK kinase (MAPKKK or MEKK) that activates a MAP/ERK kinase (MEK or MAPKK). This stimulates a phosphorylation-dependent increase in the activity of the MAP kinase. Upon activation, MAPKs phosphorylate a variety of intracellular targets including transcription factors, transcription adaptor proteins, membrane and cytoplasmic substrates as well as other protein kinases.
At least three parallel MAPK pathways exist in humans. The extracellular signal-regulated protein kinase (ERK) pathway primarily transmits mitogenic and differentiation stimuli, while the c-Jun N-terminal kinase (JNK) and p38 pathways predominantly transmit stress and cytokine stimuli. c-Myc, an ERK substrate, is a transcription factor that regulates cell growth and differentiation, glycolysis and apoptosis. Deregulation of c-Myc has been implicated in the origin of diverse human cancers. Elk-1 is a member of the ternary complex factor (TCF) sub-family of the ETS domain family. Elk-1 can be stimulated by all three MAPK pathways, and its main function is the regulation of the activity of the c-Fos promoter in response to extracellular stimuli. MEF2, a member of the MADS box family, is mainly involved in muscle differentiation, but also plays roles in muscle hypertrophy, neuronal survival and T-cell apoptosis. MEF2 is activated by both the p38 and ERK5 pathways.1 STAT1 is involved in activation of IFNα and γ genes, and is activated by p38 and JNK pathways. c-Jun is a member of the activator protein-1 (AP-1) family and is activated by both ERK1/2 and JNK pathways.2 AP-1 members play roles in the expression of genes involved in proliferation and cell cycle progression. ATF-2 is a member of the ATF/CREB family that binds to the cAMP response element (CRE). ATF-2 is activated by ERK1/2, JNK and p38.
Storage instructionsPlease refer to protocols.
Components 1 x 96 tests 5 x 96 tests 10X Antibody Binding Buffer 1 x 2.2ml 1 x 11ml 10X Wash Buffer 1 x 22ml 1 x 110ml 96-well MEF2 assay plate 1 unit 5 units Anti-rabbit HRP-conjugated IgG (0.25 μg/μL) 1 x 11µl 1 x 55µl Binding Buffer 1 x 10ml 1 x 50ml C2C12 nuclear extract (2.5 μg/μL) 1 x 40µl 1 x 200µl Developing Solution 1 x 11ml 1 x 55ml Dithiothreitol (DTT) (1 M) 1 x 100µl 1 x 500µl Lysis Buffer 1 x 10ml 1 x 50ml MEF2 antibody 1 x 11µl 1 x 55µl MEF2 mutated oligonucleotide (10 pmol/μL) 1 x 100µl 1 x 500µl MEF2 wild-type oligonucleotide (10 pmol/μL) 1 x 100µl 1 x 500µl Plate sealer 1 unit 5 units Protease Inhibitor Cocktail 1 x 100µl 1 x 500µl Stop Solution 1 x 11ml 1 x 55ml
RelevanceTranscriptional activator which binds specifically to the MEF2 element, 5'-YTA[AT]4TAR-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. Associates with chromatin to the ZNF16 promoter.
- Myocyte-specific enhancer factor 2A
- Serum response factor-like protein 1
ab207213 has not yet been referenced specifically in any publications.