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.

LanguageEnglish
Article number1900027
Pages1-3
Number of pages3
JournalProteomics
Volume19
Issue number9
Early online date13 Mar 2019
DOIs
Publication statusPublished - May 2019

Fingerprint

Immunoprecipitation
Proteomics
Proteins
Staphylococcal Protein A
Throughput
Antibodies

Keywords

  • 96-well plate
  • immunoprecipitation
  • magnetic beads
  • proteomics
  • sample preparation

Cite this

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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",
author = "Gonzalez-Lozano, {Miguel A.} and Frank Koopmans and Iryna Paliukhovich and Smit, {August B.} and Li, {Ka Wan}",
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A Fast and Economical Sample Preparation Protocol for Interaction Proteomics Analysis. / Gonzalez-Lozano, Miguel A.; Koopmans, Frank; Paliukhovich, Iryna; Smit, August B.; Li, Ka Wan.

In: Proteomics, Vol. 19, No. 9, 1900027, 05.2019, p. 1-3.

Research output: Contribution to JournalArticleAcademicpeer-review

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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

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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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