The Nrf2 (Acetyl-Lys599) Antibody Colorimetric Cell-Based ELISA Kit is a convenient, lysate-free, high throughput and sensitive assay kit that can detect Nrf2 (Acetyl-Lys599) Antibody protein expression profile in cells. The kit can be used for measuring the relative amounts of Nrf2 (Acetyl-Lys599) Antibody in cultured cells as well as screening for the effects that variuos treatments, inhibitors (ie siRNA or chemicals), or activators have on Nrf2 (Acetyl-Lys599) Antibody.
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 1)Hela cell, 2)Hela cells knockdown by NRF2-siRNA (F:CAUUGAUGUUUCUGAUCUATT,R:UAGAUCAGAAACAUCAAUGTT), (Green) primary antibody was diluted at 1:1000, 4°over night, Dylight 800 secondary antibody(Immunoway:RS23920)was diluted at 1:10000, 37° 1hour. (Red) GAPDH Monoclonal Antibody(5B7) (Immunoway:YM3029) antibody was diluted at 1:5000 as loading control, 4° over night, Dylight 680 secondary antibody(Immunoway:RS23710)was diluted at 1:10000, 37° 1hour.
Western Blot analysis of A549 cells using Acetyl-Nrf2 (K599) Polyclonal Antibody. Antibody was diluted at 1:1000. Secondary antibody(catalogNo.:SA0002) was diluted at 1:20000