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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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Updated: Jun 20, 2026

Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions

Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions

Published on: January 7, 2019

High quality protein microarray using in situ protein purification.

Keehwan Kwon¹, Carissa Grose, Rembert Pieper

¹Pathogen Functional Genomics Resource Center, J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, Maryland 20850, USA. kkwon@jcvi.org

BMC Biotechnology

|August 25, 2009

Summary

This summary is machine-generated.

Optimized in situ purification of His-tagged proteins on microarray slides enhances protein purity and enables cost-effective, high-density protein microarray generation for various applications.

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Published on: January 7, 2019

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High-throughput Protein Expression Generator Using a Microfluidic Platform

High-throughput Protein Expression Generator Using a Microfluidic Platform

Published on: August 23, 2012

Area of Science:

Biotechnology
Proteomics
Molecular Biology

Background:

High-throughput protein expression and microarray technologies are advancing.
Hexa-histidine (His-tagged) fusion proteins are commonly used for expression and purification via immobilized metal ion affinity chromatography (IMAC).
The quality of in situ His-tagged protein purification on slides requires systematic evaluation.

Purpose of the Study:

To establish and evaluate methods for determining the purification level of His-tagged proteins on metal chelate-modified slide surfaces.
To optimize in situ purification for cost-effective generation of high-quality, high-density protein microarrays.

Main Methods:

Examined two slide surfaces: chelated Cu2+ slides with polyethylene glycol (PEG) coating and chelated Ni2+ slides without PEG.
Optimized a wash buffer (PBST) for improved protein purity.
Utilized Escherichia coli cell lysates expressing 90 recombinant Streptococcus pneumoniae proteins.

Main Results:

PEG-coated chelated Cu2+ slides consistently yielded higher recombinant protein purities.
An optimized PBST wash buffer further enhanced protein purity.
Out of 90 expressed proteins, 73 were successfully immobilized, and 66 were in situ purified with >90% purity.
Identified antigens using specific antibodies and human antiserum, demonstrating immunoproteomics profiling capabilities.

Conclusions:

An optimized platform for in situ protein purification on microarray slides using His-tagged proteins is valuable for screening protein functions and interactions.
This method is suitable for immunoproteomics, complementing non-recombinant approaches for bacterial antigen discovery.