RIP-Assay Kit is optimized for performing the RIP-Chip process. In the RIP-Assay protocol, mRNP complexes are isolated from cell extracts by immunoprecipitation with RIP-Certified Anti-RBP Antibodies provided from MBL. mRNAs are isolated from mRNPs using guanidine hydrochloride. Thus, RIP-Assay Kit does not contain phenol or chloroform, allowing safe isolation of “high-quality RNA” from RNP complexes without degradation. Once purified, the RNAs present in the complex are analyzed to identify the target mRNAs using various molecular biology tools such as RT-PCR, gene expression analysis based on microarray technology (Chip analysis), or sequencing.

The major advantage of RIP-Assay Kit over most other omics approaches is that the majority of the RNAs identified exhibit structural and functional relationships. Structurally, the mRNAs in a complex contain common binding motifs for the RNA binding protein employed. Functionally, the RNAs identified generally share a common RNA regulatory network as they tend to be co-localized by the RNA binding protein which may determine how they are utilized in the cell. A second advantage of RIP-Assay Kit over traditional RNA isolation and analysis methods is that the fractionation procedure effectively concentrates the RNA species bound to a specific binding protein enabling small changes in levels of low-abundance RNAs to be detected with a greatly increased signal-to-noise ratio. When performed according to the supplied protocol, another advantage of RIP-Assay Kit is that RNA reassociation is minimized and RNAs contained in the RNP complex of interest are abundantly recovered.