Anti-MAP Kinase, Non-Phosphorylated ERK antibody, Mouse monoclonal

Mitogen-activated protein kinase (MAPK) superfamily of enzymes is involved in widespread signalling pathways. Members of this family include the ERK1/2 (extracellular signal-regulated protein kinase, also termed p42/p44 MAPK), JNK and p38 MAPK subfamilies. These are the terminal enzymes in a signalling cascade where each kinase phosphorylates and activates the next member in the sequence. Phosphorylation of both tyrosine and threonine is essential for the full activation of all MAPKs. Several kinases participate in activation of the ERK cascade. This cascade is initiated by the small G protein Ras, which upon stimulation causes activation Raf1 kinase. Raf1 continues the transmission by activating MEK. Activated MEK appears to be the only kinase capable of specifically phosphorylating and activating ERK. ERK appears to be an important regulatory molecule, which by can phosphorylate regulatory targets in the cytosol (phospholipase A2, PLA2), translocated into and phosphorylate substrates in the nucleus (ELK1). The activation of ERK cascade mediates and regulates the signal transduction pathways in response to stress, mitogenic signals and is important in development and differentiation, learning, memory and survival.

The antibody reacts specifically with the non-phosphorylated, non-activated form of MAP kinase (ERK-1 and ERK-2). Weak cross-reaction is observed with monophosphorylated (threonine or tyrosine), but not with diphosphorylated peptides of MAPK. The antibody does not recognize JNK or p38 MAPK. The epitope recognized by the antibody contains non-phosphorylated threonine 183 and tyrosine 185 which resides within the ERK-activation loop (e.g., amino acids 178-188 in ERK-2).

synthetic peptide HTGFLTEYVAT, corresponding to the non-phosphorylated form of ERK-activation loop.

A working concentration of 5-25 μg/mL may be used for detection by immunoblotting in rat brain extract. The antibody has also been used at working dilution of 1:50 for MAPK quantification by flow cytometry in monocyte-derived macrophages (MDM) derived from Holstein cattle.

Solution in phosphate buffered saline, pH 7.4, containing 15 mM sodium azide.

Prepared from a culture supernatant of bioreactor grown hybridoma

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