The QuantiSir™ Specific Gene Knockdown Quantification Kit For Signal Transduction series, offering a flexible choice of different kits used for measuring the knockdown of 50 common genes related to signal transduction, is part of Epigentek's advanced gene knockdown assay system to quantify gene knockdown induced by siRNA or antisense oligonucleotide at the protein level in cultured cells or tissues. The kit offers a more highly convenient process than traditional methods such as Northern blot, quantitative RT-PCR, and Western Blot. The kit addresses and corrects traditional problems such as the decrease in the amount of a specific mRNA not typically correlating well with protein levels present in the cell, and the lack of ability to discriminate between samples in which the differences in protein levels are minimal. The kit has the following advantages:
Principle & ProcedureThe QuantiSir™ Specific Gene Knockdown Quantification Kit For Signal Transduction is specifically designed for quantifying gene knockdown induced by siRNA or antisense oligonucleotide at the protein level in the cultured cells or tissues. In the assay, the cell lysates containing the targeted protein are stably spotted on the specifically treated microwells with a unique protein capture buffer. The spotted protein can then be recognized with the target-specific antibody and colorimetrically measured through detection antibody-chromogen reaction system.
Fig. 1. Schematic procedure for using the QuantiSir Specific Gene Knockdown Quantification Kit For Signal Transduction.
Fig. 2. Quantification of HDAC1 knockdown. MCF-7 cells were treated or untreated with HDAC1 siRNA. Protein extracts were prepared and used for detection of HDAC1 protein level.
Q1 (Extraction Buffer)Q2 (10X Wash Buffer)Q3 (Protein Capture Buffer)Q4 (Blocking Buffer)Q5 (Antibody Buffer)Q6 (Developing Solution)Q7 (Stop Solution)GAPDH Control Antibody*Capture Antibody*Detection Antibody*8-Well Assay Strips (with Frame)User Guide* For maximum recovery of the products, centrifuge the original vial after thawing prior to opening the cap.