Cytokine Detection by Antibody-based Proximity Ligation

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2004 May 25

PMID 15155907

Citations 133

Authors

Mats Gullberg

Sigrun M Gustafsdottir

Edith Schallmeiner

Jonas Jarvius

Mattias Bjarnegard

Christer Betsholtz

Ulf Landegren

Simon Fredriksson

Affiliations

Soon will be listed here.

Abstract

Efficient and precise detection techniques, along with extensive repertoires of specific binding reagents, will be needed to meet the challenges of proteome analyses. The recently established proximity ligation mechanism enables sensitive high-capacity protein measurements by converting the detection of specific proteins to the analysis of DNA sequences. Proximity probes containing oligonucleotide extensions are designed to bind pairwise to target proteins and to form amplifiable tag sequences by ligation when brought in proximity. In our previous report, both the ligatable arms and the protein binders were DNA molecules. We now generalize the method by providing simple and convenient protocols to convert any polyclonal antibodies or matched pair of monoclonal antibodies to proximity probe sets through the attachment of oligonucleotide sequences. Sufficient reagent for >100,000 proximity ligation assays can be prepared from 1 microg of antibody. The technique is applied to measure cytokines in a homogenous test format with femtomolar detection sensitivities in 1-microl samples, and we exemplify its utility in situations when only minute sample amounts are available.

Citing Articles

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Use of Proximity Ligation Assay to Study Lymphoid Malignancies.

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