Description
Key facts
Detection method:Colorimetric,
Sample types:Nuclear Extracts,
Reacts with:Mouse, Human,
Assay type:Semi-quantitative,
Sensitivity:= 500 ng/well,
Assay time:3h 30m,
Assay Platform:Microplate reader
Product details:
MAPK (ATF2, c-Jun, c-Myc, MEF2, STAT1alpha) Transcription Factor Assay Kit (Colorimetric) (ab207212) is a high throughput assay to quantify activation of MAPK regulated transcription factors. 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 MAPK consensus binding site has been immobilized onto a 96-well plate. Active MAPK present in the nuclear extract specifically binds to the oligonucleotide. MAPK is detected by a primary antibody that recognizes an epitope of MAPK 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 human MAPK and mice c-Myc, c Jund and Jun D.
Key performance and benefits:
Assay time: 3.5 hours (cell extracts preparation not included).
Detection limit: < 0.5 μg nuclear extract/well.
Detection range: 0.5 – 5 μg of nuclear extract/well for c-Myc, MEF2 and STAT1; 1 – 10 μg nuclear extract/well for ATF-2 and c-Jun.
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.
Properties and Storage Information:
Shipped at conditions-Dry Ice, Appropriate short-term storage conditions-Multi, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols
