RayBio® Phospho-AKT(Ser473) Quantitative ELISA kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in Human cell lysates. By determining phosphorylated AKT protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human phospho-AKT. An anti-pan AKT antibody has been coated onto a 96-well plate. Standard and Samples are pipetted into the wells and AKT present in a sample is bound to the wells by the immobilized antibody. The wells are washed and rabbit anti-phospho-AKT(Ser473) antibody is used to detect phosphorylated AKT. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of AKT(Ser473) bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450nm.

Upon receipt, the kit should be stored at -20 °C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), HRP-Streptavidin (Item G), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4 °C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge and store at -20 °C. Reconstituted Positive Control (Item K) should be stored at -70 °C.