Title |
Beads-free protein immunoprecipitation for a mass spectrometry-based interactome and posttranslational modifications analysis
|
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Published in |
Proteome Science, September 2015
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DOI | 10.1186/s12953-015-0079-0 |
Pubmed ID | |
Authors |
Michal Mikula, Tymon Rubel, Jakub Karczmarski, Malgorzata Statkiewicz, Karol Bomsztyk, Jerzy Ostrowski |
Abstract |
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. |
X Demographics
Geographical breakdown
Country | Count | As % |
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United Kingdom | 1 | 50% |
Unknown | 1 | 50% |
Demographic breakdown
Type | Count | As % |
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Members of the public | 2 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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United Kingdom | 1 | 3% |
Unknown | 38 | 97% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 15 | 38% |
Researcher | 8 | 21% |
Student > Bachelor | 6 | 15% |
Student > Master | 3 | 8% |
Professor | 2 | 5% |
Other | 2 | 5% |
Unknown | 3 | 8% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 15 | 38% |
Biochemistry, Genetics and Molecular Biology | 10 | 26% |
Medicine and Dentistry | 3 | 8% |
Neuroscience | 3 | 8% |
Immunology and Microbiology | 2 | 5% |
Other | 2 | 5% |
Unknown | 4 | 10% |