RNA Immunoprecipitation Service
  • OVERVIEW

RNA Immunoprecipitation Service


The relevance of RNA-protein interactions in modulating mRNA and noncoding RNA function is increasingly appreciated and several methods have been recently developed to map them. The RNA immunoprecipitation (RIP) is a powerful method to study the physical association between individual proteins and RNA molecules in vivo.

Several variants of this technique exist and can be divided into two main classes: native and cross-linked RNA immunoprecipitation. The native RIP allows to reveal the identity of RNAs directly bound by the protein and their abundance in the immunoprecipitated sample, while cross-linked RIP leads to precisely map the direct and indirect binding site of the RBP of interest to the RNA molecule. Live cells are treated with formaldehyde to generate protein-RNA cross-links between proximal molecules. Following immunoprecipitation of a protein of interest and cross-link reversal, associated RNAs can be recovered, characterized, and quantitated by reverse transcriptase polymerase chain reaction (RT-PCR). Protein association with specific RNA regions can be performed under a variety of conditions (e.g., different environmental and cell-cycle states) and/or in mutant strains. Furthermore, because formaldehyde inactivates cellular enzymes essentially immediately upon addition to cells, RIP provides snapshots of protein-RNA interactions at specific time points and hence is useful for kinetic analyses of events occurring on RNA in vivo.


Our Solutions

-Protein/RNA interaction analysis(q-PCR)

-RNA/RNA interaction analysis(q-PCR)

-Protein/RNA interaction analysis(sequencing)

-RNA/RNA interaction analysis(sequencing)


Workflow of Our Service

-Vector construction and transient expression

-Cell lysate preparation

-Magnetic beads preparation

-RNA-binding protein immunoprecipitation

-RNA purification

-RNA reverse transcription and cDNA verification (please inquire if sequencing analysis is required)

-Data analysis and report writing


Sample Requirements

-Target protein antibody (suitable for IP application)

-Cell/tissue samples to be tested

-Target gene, target protein information


Deliverables

-Raw data

-Project report

RNA Immunoprecipitation Service


The relevance of RNA-protein interactions in modulating mRNA and noncoding RNA function is increasingly appreciated and several methods have been recently developed to map them. The RNA immunoprecipitation (RIP) is a powerful method to study the physical association between individual proteins and RNA molecules in vivo.

Several variants of this technique exist and can be divided into two main classes: native and cross-linked RNA immunoprecipitation. The native RIP allows to reveal the identity of RNAs directly bound by the protein and their abundance in the immunoprecipitated sample, while cross-linked RIP leads to precisely map the direct and indirect binding site of the RBP of interest to the RNA molecule. Live cells are treated with formaldehyde to generate protein-RNA cross-links between proximal molecules. Following immunoprecipitation of a protein of interest and cross-link reversal, associated RNAs can be recovered, characterized, and quantitated by reverse transcriptase polymerase chain reaction (RT-PCR). Protein association with specific RNA regions can be performed under a variety of conditions (e.g., different environmental and cell-cycle states) and/or in mutant strains. Furthermore, because formaldehyde inactivates cellular enzymes essentially immediately upon addition to cells, RIP provides snapshots of protein-RNA interactions at specific time points and hence is useful for kinetic analyses of events occurring on RNA in vivo.


Our Solutions

-Protein/RNA interaction analysis(q-PCR)

-RNA/RNA interaction analysis(q-PCR)

-Protein/RNA interaction analysis(sequencing)

-RNA/RNA interaction analysis(sequencing)


Workflow of Our Service

-Vector construction and transient expression

-Cell lysate preparation

-Magnetic beads preparation

-RNA-binding protein immunoprecipitation

-RNA purification

-RNA reverse transcription and cDNA verification (please inquire if sequencing analysis is required)

-Data analysis and report writing


Sample Requirements

-Target protein antibody (suitable for IP application)

-Cell/tissue samples to be tested

-Target gene, target protein information


Deliverables

-Raw data

-Project report

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