71378  NoShift™ Transcription Factor Assay Kit Plus NucBuster™

An easy and rapid alternative to EMSA

The NoShift™ Transcription Factor Assay Kit Plus NucBuster™ incorporates the NoShift™ Transcription Factor Assay Kit (Cat. No. 71377) with NucBuster™ Protein Extraction Reagent for convenient production of nuclear protein extracts. The NoShift™ Transcription Factor Assay Kit is a microassay plate–based method for analysis of DNA-binding proteins that alleviates many of the problems associated with gel shift assays. The gel shift, or electrophoretic mobility shift assay (EMSA), is a method for analyzing interactions between proteins and DNA. This technique, first published in 1981 (Fried 1981, Garner 1981), is based on the reduced electrophoretic mobility of protein–DNA complexes compared with that of DNA alone under nondenaturing conditions. The traditional gel shift assay is tedious and time consuming, requires radio-labeled probe, and often requires extensive optimization. The 96-well NoShift plate format is versatile and enables simultaneous analysis of multiple binding factors in less than five hours without the use of radioisotopes. A biotinylated oligonucleotide is mixed with a protein sample and the protein–DNA complex is captured on streptavidin plates. The bound transcription factor is detected with a specific antibody followed by a second antibody–HRP conjugate and chromogenic substrate.

Each NoShift Transcription Factor Assay Kit comes with one 96-strip-well streptavidin-coated plate, binding reaction buffer, antibody dilution buffer, plate wash buffer, and TMB substrate and provides enough reagents for 100 reactions. Also available is the NoShift Transcription Factor Assay Kit Plus Nucbuster™ which includes a NucBuster™ Protein Extraction Kit developed for rapid extraction of nuclear protein. The NoShift Kit is designed for use with oligonucleotides defining a protein binding consensus sequence provided by the purchaser and an antibody to a DNA binding protein.