QuantiSir General Gene Knockdown Quantification Kit

The QuantiSir™ General Gene Knockdown Quantification Kit 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:

• Quick and efficient procedure -- completion of entire assay requires only 4 hours.
• Innovative colorimetric assay without the need for radioactivity, electrophoresis, or chromatography.
• The internal control is conveniently included in the kits to correct for the variations in the cell number or protein concentrations.
• Strip microwell format makes the assay flexible: manual or high throughput.
• Simple, reliable, and consistent assay conditions.

Principle & Procedure
The QuantiSir™ General Gene Knockdown Quantification Kit 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 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. GAPDH expression as the internal control can be used for normalizing the effect of target gene knockdown.

 
Fig. 1. Schematic procedure for using the QuantiSir General Gene Knockdown Quantification Kit.

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.