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Immunoprecipitation

Immunoprecipitation

Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...

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Related Experiment Video

Updated: Sep 8, 2025

Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution

Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution

Published on: January 16, 2017

R-Loop Immunoprecipitation: A Method to Detect R-Loop Interacting Factors.

Chiara Beghè¹, Natalia Gromak²

¹Sir William Dunn School of Pathology, University of Oxford, Oxford, UK.

Methods in Molecular Biology (Clifton, N.J.)

|June 15, 2022

Summary

This study presents a method to detect R-loop interactors, crucial for understanding genome stability and diseases like cancer. The protocol uses S9.6 antibody affinity purification, aiding in the discovery of new R-loop binding factors.

Keywords:

APOBEC3B Affinity purification DHX9 Immunoprecipitation R-loop R-loop interactors RNA/DNA hybrids S9.6 antibody Slot Blot

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Identification of Footprints of RNA:Protein Complexes via RNA Immunoprecipitation in Tandem Followed by Sequencing RIPiT-Seq

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Published on: July 10, 2019

Method for the Isolation and Identification of mRNAs, microRNAs and Protein Components of Ribonucleoprotein Complexes from Cell Extracts using RIP-Chip

Method for the Isolation and Identification of mRNAs, microRNAs and Protein Components of Ribonucleoprotein Complexes from Cell Extracts using RIP-Chip

Published on: September 29, 2012

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Last Updated: Sep 8, 2025

Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution

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Published on: January 16, 2017

Identification of Footprints of RNA:Protein Complexes via RNA Immunoprecipitation in Tandem Followed by Sequencing RIPiT-Seq

Identification of Footprints of RNA:Protein Complexes via RNA Immunoprecipitation in Tandem Followed by Sequencing RIPiT-Seq

Published on: July 10, 2019

Method for the Isolation and Identification of mRNAs, microRNAs and Protein Components of Ribonucleoprotein Complexes from Cell Extracts using RIP-Chip

Method for the Isolation and Identification of mRNAs, microRNAs and Protein Components of Ribonucleoprotein Complexes from Cell Extracts using RIP-Chip

Published on: September 29, 2012

Area of Science:

Molecular Biology
Genetics
Biochemistry

Background:

R-loops are non-B-DNA structures formed by an RNA/DNA hybrid and a displaced single-stranded DNA during transcription.
Aberrant R-loop levels are linked to DNA damage, genome instability, and diseases such as cancer and neurodegeneration.

Purpose of the Study:

To describe a protocol for detecting R-loop interacting factors.
To enable the identification and characterization of novel proteins involved in R-loop biology.

Main Methods:

Affinity purification using the S9.6 antibody, which specifically targets RNA/DNA hybrids.
Detection of bound factors via Western blotting or mass spectrometry.
Specificity verification using synthetic competitors and RNase H treatment.

Main Results:

Successful identification of R-loop binding factors.
Validation of the specificity of identified factors through rigorous controls.
Establishment of a method to discover new R-loop interactors.

Conclusions:

The described protocol provides a robust method for identifying R-loop interacting proteins.
This approach serves as a valuable resource for investigating the roles of R-loops in cellular processes and disease pathogenesis.
Understanding R-loop interactions is key to developing therapeutic strategies for R-loop-associated diseases.