The CellSimple™ Akt and MAPK Activation Antibody Assay Kit is designed for use with the CellSimple™ Cell Analyzer. The kit contains Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (197G2) Rabbit mAb (PE Conjugate) #14095 and Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) #4071. The kit allows for the quick and easy measurement of relative phosphorylation levels of ERK 1/2 MAPK and Akt under given experimental conditions.
Antibodies provided in the CellSimple™ Akt and MAPK Activation Antibody Assay Kit detect endogenous levels of their respective target. Additionally the Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (197G2) Rabbit mAb (PE Conjugate) does not cross-react with the corresponding phosphorylated residues of either JNK/SAPK or p38 MAP kinase.
The CellSimple™ Cell Analyzer is a benchtop instrument that utilizes a disposable thin-film cassette and a combination of a 488 nm laser, two photomultiplier tubes (525/45 nm and 561 nm LP filters), Coulter Principle-based cell measurements, and on-board software to provide easy-to-run applications and data analysis. Data acquisition occurs within approximately 10 seconds per test. The instrument relies on disposable cassettes for sample handling, which alleviates the need for flow cell cleaning and fluidics maintenance and the instrument is small enough to be portable between the lab bench and the hood. Applications include quantitative assessments of cell viability, apoptosis, other labeled antibody markers and single and multiplexed bead-based assays for protein and cellular analysis.
The MAP Kinase (MAPK) and the PI-3 Kinase/Akt pathways are two of the most well characterized and widely studied signaling modules which transmit signals emanating from the extracellular environment to alter cell behavior. They are involved in a broad range of cellular processes such as cell proliferation, survival, differentiation, and motility. The ERK 1 and 2 (from the MAPK pathway) and Akt protein kinases are two quintessential components serving as integration hubs in their respective pathways. ERK 1/2 protein kinases are activated by dual phosphorylation on Thr202 and Tyr204 from the the dual specificity kinases MEK1 and MEK2. The Akt protein kinase is activated by phosphorylation on Ser473 and Thr308 from the TORC2 complex and PDK1, respectively. The relative levels of these phosphorylations are reliable proxies of activation levels of these kinases. The knowledge about the levels of activation of these kinases is instructive in a broad range of experimental settings making these two molecular events some of the most popular and sought after assay endpoints.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
CellSimple is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
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