Dataset Information

A method to characterize antibody selectivity immunoprecipitation, part 2

ABSTRACT: Antibodies are used in multiple cell biology applications but there are methods to assess antibody quality, riskingdata integrity and repr developed a standard operating procedure using HEK293 cells to score and antibodies for suitability in immunoprecipitation. This method, validated in in fourindependent laboratories, was developed using 1124 novel recomb for152 chromatin-related human proteins. We used mass spectrometry abundanceof all the proteinsin eachimmunoprecipitateand compared spectral abundance factors from the target antigen with those of all Antibodies for which the targetantigen or a member of its known protein most abundant protein were classified as “IP gold standard”. This m quantitative outputs that can be stored and archived in public databases, step towards a platform for community benchmarking of antibody quality.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Hongbo Guo

LAB HEAD: Jack Greenblatt

PROVIDER: PXD002534 | Pride | 2015-07-15

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	FINV_uniprot_sprot_human_BSA_TRY_BCN_GFP_TEV_062911.fasta	Fasta
	GR_121213_10_OT3_CHD2_B005_1_HEK293.mzXML	Mzxml
	GR_121213_11_OT3_CHD2_B006_1_HEK293.mzXML	Mzxml
	GR_121213_12_OT3_CHD9_B004_1_HEK293.mzXML	Mzxml
	GR_121213_13_OT3_CHD4_B001_1_HEK293.mzXML	Mzxml

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Publications

Assessment of a method to characterize antibody selectivity and specificity for use in immunoprecipitation.

Marcon Edyta E Jain Harshika H Bhattacharya Anandi A Guo Hongbo H Phanse Sadhna S Pu Shuye S Byram Gregory G Collins Ben C BC Dowdell Evan E Fenner Maria M Guo Xinghua X Hutchinson Ashley A Kennedy Jacob J JJ Krastins Bryan B Larsen Brett B Lin Zhen-Yuan ZY Lopez Mary F MF Loppnau Peter P Miersch Shane S Nguyen Tin T Olsen Jonathan B JB Paduch Marcin M Ravichandran Mani M Seitova Alma A Vadali Gouri G Vogelsang Maryann S MS Whiteaker Jeffrey R JR Zhong Guoqing G Zhong Nan N Zhao Lei L Aebersold Ruedi R Arrowsmith Cheryl H CH Emili Andrew A Frappier Lori L Gingras Anne-Claude AC Gstaiger Matthias M Paulovich Amanda G AG Koide Shohei S Kossiakoff Anthony A AA Sidhu Sachdev S SS Wodak Shoshana J SJ Gräslund Susanne S Greenblatt Jack F JF Edwards Aled M AM

Nature methods 20150629 8

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 fro ...[more]

PMID: 26121405

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