Description: The InstantOne™ ELISA is specifically engineered for accurate measurement of phosphorylated ERK 1/2 (human, mouse, hamster), AKT 1/2/3 (human, mouse, hamster), and p70 S6K (human, mouse) in cell lysates. Detection of these phosphorylated molecules in rat is also expected. The InstantOne™ ELISA kit allows for fast analysis of samples in approximately one hour. All reagents used in a traditional sandwich ELISA are added in solution to a plate followed by a wash step and detection with the TMB colorimetric substrate.
Extracellular signal-regulated kinase (ERK) is the founding member and a key component of the classical Mitogen-Activated Protein Kinase (MAPK) pathway. MEK activates ERK 1/2 by phosphorylation of its TxY motifs in response to a variety of stimuli, including growth factors and cytokines.
AKT, which exists as multiple isoforms, is one of the principle kinases activated by phosphoinositide 3-kinase (PI3K). These lipid second messengers bind to the pleckstrin homology domain of AKT to promote its translocation to the plasma membrane for activation via phosphorylation at Thr308 and Ser473 by PDK1 and the mTOR TORC2 complex, respectively. Phosphorylation at both these sites is required for full activation of AKT Ser/Thr kinase activity.
p70 S6 Kinase (p70S6K) is a member of the ribosomal S6 kinase family of Ser/Thr kinases. p70 S6K activity is controlled by multiple signaling pathways, including the MAPK, phosphoinositide-3 kinase (PI3K), and mTOR pathways. This regulation results in the phosphorylation of multiple sites of p70 S6K, including Thr389 within the linker domain, which is required for full activation. Activated p70 S6K phosphorylates several residues on the S6 ribosomal protein, thereby leading to increased protein synthesis.
This kit has been developed to recognize the following phosphorylation sites:
ERK 1/2/3 (Thr202/Tyr204, Thr185/Tyr187)
AKT 1/2/3 (Ser473)
p70 S6K (Thr389)