The p70 S6 Kinase (Phospho-Thr389) Colorimetric Cell-Based ELISA Kit is a convenient, lysate-free, high throughput and sensitive assay kit that can detect p70 S6 Kinase (Phospho-Thr389) protein expression profile in cells. The kit can be used for measuring the relative amounts of p70 S6 Kinase (Phospho-Thr389) in cultured cells as well as screening for the effects that variuos treatments, inhibitors (ie siRNA or chemicals), or activators have on p70 S6 Kinase (Phospho-Thr389).
This kit contains Crystal Violet Staining normalization protocol. It provides a means to normalize Absorbance 450 nm of primary antibodies to cell density of the cells in the same well. This provides an accurate account for stimulations and cell densities during the assay. By comparing the proportional values obtained from the experimental wells to those of the control wells (e.g. untreated cells), it is possible to determine the extent to which the stimulant(s) have affected the expression levels.
Western blot analysis of lysates from Hela cells, treated with LPS 100ng/ml 30', using total P70 S6 Kinase Antibody(catalog: YT3555) and P70 S6 Kinase (phospho T389) antibody(catalog: YP1427).(Green) primary antibody was diluted at 1:1000, 4°over night, secondary antibody(cat:RS23920)was diluted at 1:10000, 37° 1hour.