A Fast and Economical Sample Preparation Protocol for Interaction Proteomics Analysis

Miguel A. Gonzalez-Lozano; Frank Koopmans; Iryna Paliukhovich; August B. Smit; Ka Wan Li

doi:10.1002/pmic.201900027

A Fast and Economical Sample Preparation Protocol for Interaction Proteomics Analysis

Miguel A. Gonzalez-Lozano, Frank Koopmans, Iryna Paliukhovich, August B. Smit, Ka Wan Li^*

^*Corresponding author for this work

Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

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.

Original language	English
Article number	1900027
Pages (from-to)	1-3
Number of pages	3
Journal	Proteomics
Volume	19
Issue number	9
Early online date	13 Mar 2019
DOIs	https://doi.org/10.1002/pmic.201900027
Publication status	Published - May 2019

Keywords

96-well plate
immunoprecipitation
magnetic beads
proteomics
sample preparation

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This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1002/pmic.201900027

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title = "A Fast and Economical Sample Preparation Protocol for Interaction Proteomics Analysis",

abstract = "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.",

keywords = "96-well plate, immunoprecipitation, magnetic beads, proteomics, sample preparation",

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T1 - A Fast and Economical Sample Preparation Protocol for Interaction Proteomics Analysis

AU - Gonzalez-Lozano, Miguel A.

AU - Koopmans, Frank

AU - Paliukhovich, Iryna

AU - Smit, August B.

AU - Li, Ka Wan

PY - 2019/5

Y1 - 2019/5

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AB - 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.

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KW - immunoprecipitation

KW - magnetic beads

KW - proteomics

KW - sample preparation

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A Fast and Economical Sample Preparation Protocol for Interaction Proteomics Analysis

Abstract

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