Beads-free protein immunoprecipitation for a mass spectrometry-based interactome and posttranslational modifications analysis.
Protein immunoprecipitation (IP) coupled with MS provides means to interrogate protein complexes and their posttranslational modifications (PTMs). In a typical protein IP assay antibodies are conjugated to protein A/G beads requiring large amounts of antibodies, tube transfers and centrifugations.As an alternative, we present Matrix-IP, beads-free microplate-based platform with surface-immobilized antibodies. ... Assay utilizes standard 96-well polypropylene PCR plates that are laboratory-fabricated with UV-C light and then protein A/G coated prior to IP reaction. We demonstrate application of Matrix-IP platform in MS analysis of heterogeneous nuclear ribonucleoprotein K (hnRNP K) interactome and PTMs.Matrix-IP is time-saving, easy to use high throughput method adaptable for low sample amounts and automation.
Proteome Sci
Date: Sep. 04, 2015
PubMed ID: 26336360
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