Blocking Buffer 20X Tris Buffered Saline Surfact-Amps 20 Surfact-Amps X-100 HRP Conjugate TMB Substrate TMB Stop Solution Janus Green Whole-Cell Stain Elution Buffer Thin Plate Seal Assembly Components included only in the target-specific kits:Antibody #1 and Antibody #2
The AKT Multispecies In-Cell ELISA Colorimetric Detection Kit is a simple method for quantifying intracellular proteins in whole cells.
Principle of the method
To perform the assay, cells are first plated, treated and fixed. Expression of the protein(s) of interest is monitored in wells of a microplate using target-specific primary antibodies (see the Important Product Information section for antibodies included in each kit) and a horseradish peroxidase (HRP)-conjugated detection reagent. The kit is supplied with a whole-cell stain to control for differences in cell plating, which is important when measuring relative levels of a protein with different treatments or assessing its post-translational modification (PTM) form. After staining, the results are analyzed by normalizing the absorbance (HRP activity) values to cell number, which adjusts for the cell plating differences among the wells.
Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.
AKT (protein kinase B, PKB, RAS-alpha) is a 57kDa serine/threonine kinase that plays an important role in diverse biological responses such as regulation of metabolism, cell survival and growth by phosphorylating multiple proteins. AKT is activated by insulin, PI3K, IGF1 and various other growth and survival factors. AKT promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including forkhead transcription factors, and caspase-9. There are three mammalian isoforms of Akt: AKT1 (PKB alpha), AKT2 (PKB beta) and AKT3 (PKB gamma) with AKT2 and AKT3 being approximately 82% identical with the AKT1 isoform. Each isoform has a pleckstrin homology (PH) domain, a kinase domain and a carboxy terminal regulatory domain. AKT was originally cloned from the retrovirus AKT8, and is a key regulator of many signal transduction pathways. The AKT pathway is a major target for cancer therpeutics where AKT signaling dysfunction has implicated in many types of cancer, including cancer syndromes known as phakomatoses.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.