Reichert, M;
Bhamra, A;
Kupka, S;
Walczak, H;
(2018)
Characterization of the TNFR1-SC Using “Modified Tandem Affinity Purification” in Conjunction with Liquid Chromatography–Mass Spectrometry (LC-MS).
In: Ting, AT, (ed.)
Programmed Necrosis. Methods and Protocols.
(pp. 161-169).
Springer Nature: New York (NY), USA.
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Abstract
Mass spectrometry enables the unbiased characterization of protein complexes. The success of this approach and the amount of information that can be retrieved are highly dependent on the achieved purity of the protein complex to be analyzed. Here we describe a modified tandem affinity purification (moTAP) approach which can be used to isolate the tumor necrosis factor receptor 1 signaling complex for subsequent analysis by liquid chromatography–tandem mass spectrometry. Indeed, this approach can easily be adapted to the isolation of other membrane-bound and intracellular signaling complexes. This methodology allows for a highly sensitive analysis and characterization of complex components, including posttranslational modifications and for the identification of novel complex components.
| Type: | Book chapter |
|---|---|
| Title: | Characterization of the TNFR1-SC Using “Modified Tandem Affinity Purification” in Conjunction with Liquid Chromatography–Mass Spectrometry (LC-MS) |
| Location: | United States |
| ISBN: | 978-1-4939-8753-5 |
| ISBN-13: | 978-1-4939-8754-2 |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1007/978-1-4939-8754-2_16 |
| Additional information: | This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions. |
| Keywords: | Tandem affinity purification, Immunoprecipitation, Proteomics, Tumor necrosis factor (TNF), TNF receptor 1 signaling complex (TNFR1-SC), Liquid chromatography-tandem mass spectrometry (LC-MS/MS) |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute > Research Department of Cancer Bio |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10057619 |
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