Immunoprecipitation (IP)-based techniques are one of the key techniques to study the interaction between proteins and other macromolecules, such as Co-IP (Co-immunoprecipitation) to study protein-protein interactions, RIP (RNA binding protein immunoprecipitation assay) to study protein-RNA interactions, and ChIP (chromatin immunoprecipitation assay) to study protein-DNA interactions.
RIP technology mainly uses antibodies against target proteins to precipitate the corresponding RNA-protein complexes, which can be separated and purified for q-PCR validation or sequencing analysis of the RNA bound to the complexes. RIP is a technique to study intracellular RNA-protein binding, and its application fields include post-transcriptional regulation studies and epigenetic regulation. RNA-binding proteins (RBPs) are involved in the transcription and maturation of mRNAs, including biological processes such as processing, translocation, stabilization, degradation and translation; similarly, some mRNA molecules are regulated by ribonucleoprotein complexes (RNPs) at the post-transcriptional level ; RIP is a powerful tool to understand the dynamic processes of post-transcriptional regulatory networks and can help discover the regulatory targets of miRNAs.
RNA immunoprecipitation (RIP-seq) done by targeting RNA binding proteins
a) Use of antibodies or epitope tags to capture endogenous RBPs in the nucleus or cytoplasm.
b) Preventing the binding of non-specific RNAs.
c) Immunoprecipitation to isolate RBPs together with their bound RNAs.
d) The bound RNA sequences are identified by quantitative RT-PCR or high-throughput sequencing (RIP-Seq) methods.
RIP can reflect the interaction between RNA and protein in the natural cellular state;
Identify RNAs that directly/indirectly interact with RBPs
RIP-q-PCR verifies the proteins screened by RNA pull down-MS to avoid false positives.
Depending on your needs, Creative Proteomics can provide RIP technology services to validate RNA/protein, RNA/RNA interactions.
All you need to do is provide cells, tissues or RNA and we will do the whole process from sample processing, library preparation, sequencing to data analysis for you.
Commercial RIP kits:
We use commercial RIP kits to ensure the success rate and stability of RIP experiments.
Strict quality control:
We monitor the quality of experiments throughout to ensure high-quality data.
Specialized bioinformatics analysis:
With a strong bioinformatics team, we are able to meet all types of in-depth data analysis requirements of our clients.
Sample type: Cell, tissue or RNA.
a) Cells: 5×108
b) Tissue: 500 mg
c) RNA after IP: >1 μg, OD260/280≥1.8; OD260/230≥1.5
PCR results or original report of sequencing.
Experimental report (experimental instruments, reagents, methods, results, conclusions).