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.
The conjugated antibodies in this kit are supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibodies. Protect from light. Do not freeze. The remaining components of the kit should be stored at the recommended temperature upon receipt. All components in this kit are stable for at least 12 months when stored at the recommended temperature and left unused.
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. This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or firstname.lastname@example.org.