| Literature DB >> 26121405 |
Edyta Marcon1, Harshika Jain2, Anandi Bhattacharya2, Hongbo Guo1, Sadhna Phanse1, Shuye Pu3, Gregory Byram4, Ben C Collins5, Evan Dowdell6, Maria Fenner2, Xinghua Guo1, Ashley Hutchinson2, Jacob J Kennedy7, Bryan Krastins4, Brett Larsen8, Zhen-Yuan Lin8, Mary F Lopez4, Peter Loppnau2, Shane Miersch1, Tin Nguyen9, Jonathan B Olsen9, Marcin Paduch6, Mani Ravichandran2, Alma Seitova2, Gouri Vadali4, Maryann S Vogelsang4, Jeffrey R Whiteaker7, Guoqing Zhong1, Nan Zhong2, Lei Zhao7, Ruedi Aebersold10, Cheryl H Arrowsmith2, Andrew Emili11, Lori Frappier9, Anne-Claude Gingras12, Matthias Gstaiger13, Amanda G Paulovich7, Shohei Koide6, Anthony A Kossiakoff6, Sachdev S Sidhu1, Shoshana J Wodak14, Susanne Gräslund2, Jack F Greenblatt11, Aled M Edwards2.
Abstract
Antibodies are used in multiple cell biology applications, but there are no standardized methods to assess antibody quality-an absence that risks data integrity and reproducibility. We describe a mass spectrometry-based standard operating procedure for scoring immunoprecipitation antibody quality. We quantified the abundance of all the proteins in immunoprecipitates of 1,124 new recombinant antibodies for 152 chromatin-related human proteins by comparing normalized spectral abundance factors from the target antigen with those of all other proteins. We validated the performance of the standard operating procedure in blinded studies in five independent laboratories. Antibodies for which the target antigen or a member of its known protein complex was the most abundant protein were classified as 'IP gold standard'. This method generates quantitative outputs that can be stored and archived in public databases, and it represents a step toward a platform for community benchmarking of antibody quality.Entities:
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Year: 2015 PMID： 26121405 DOI： 10.1038/nmeth.3472
Source DB: PubMed Journal: Nat Methods ISSN： 1548-7091 Impact factor: 28.547