RIP-Assay Kit

Post-transcriptional regulation of gene expression is a ribonucleoprotein (RNP)-driven process, which involves RNA binding proteins (RBPs) and noncoding RNAs that regulate splicing, nuclear export, subcellular localization, mRNA stability and translation. This area has recently become the focus of many research groups and progress is being made using the yeast, Saccharomyces cerevisiae and various types of mammalian cell systems. Those observations have confirmed the posttranscriptional RNA operon concept in which mRNAs that encode functionally related proteins are coordinately regulated during cellular processes such as proliferation, differentiation or drug treatment. For example, mRNAs encoding proteins that function in a particular cell process or pathway can be found within a unique mRNP complex, which consists of mRNA and RNP. This provides valuable information regarding not only known components of a particular process or pathway, but importantly, leads to the identification of novel components representing potential therapeutic targets and biomarkers. In addition to those targets identified by pathway expansion, the specific RBPs regulating RNA functions may be potential therapeutic targets in their own right. In order to understand posttranscriptional control of gene expression, RIP-Chip technologies that allow the isolation and identification of mRNAs, microRNAs and protein components of RNP complexes from cell extracts using antibodies to RBPs and microarrays have been developed.

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 RBP Antibodies provided from MBLI. 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.

Lysis buffer, Wash buffer, Normal Rabbit IgG, High-Salt Solution, Solution I, Solution II, Solution III, Solution IV

2 years

RNP Complex