A Fast and Economical Sample Preparation Protocol for Interaction Proteomics Analysis.
96-well plate
immunoprecipitation
magnetic beads
proteomics
sample preparation
Journal
Proteomics
ISSN: 1615-9861
Titre abrégé: Proteomics
Pays: Germany
ID NLM: 101092707
Informations de publication
Date de publication:
05 2019
05 2019
Historique:
received:
16
01
2019
revised:
22
02
2019
pubmed:
14
3
2019
medline:
30
4
2020
entrez:
14
3
2019
Statut:
ppublish
Résumé
A simple and fast immunoprecipitation (IP) protocol is designed with the sample preparation incorporated, applicable to both low and high throughput. This new protocol combines two procedures based on magnetic beads in 96-well plate format. Protein complexes are captured by antibodies and magnetic beads conjugated with protein A. Proteins are washed and on-bead digested by using Single-Pot solid-phase sample preparation (SP3). The whole IP-SP3 approach can be completed in one day, which is considerably faster compared to the classical approach. No major quantitative differences are found between SP3 and FASP (filter-aided sample preparation) or a longer incubation protocol. Taken together, the IP-SP3 protocol is a fast and economical approach easily applicable for large-scale protein interactome analysis.
Identifiants
pubmed: 30864274
doi: 10.1002/pmic.201900027
doi:
Substances chimiques
Antibodies
0
Multiprotein Complexes
0
Proteome
0
Staphylococcal Protein A
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e1900027Informations de copyright
© 2019 The Authors. Proteomics published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.