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Related Concept Videos

Sample Preparation for Analysis: Overview01:21

Sample Preparation for Analysis: Overview

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Sample preparation is an essential step in the analytical process. It involves preparing a sample so that it can be analyzed accurately. The goal is to extract the analyte, the substance you want to measure, from the sample while removing any components that may interfere with the analysis. Sample preparation techniques vary depending on the physical state of the sample.
Bulk or large solid samples are typically reduced in size using grinding, crushing, or milling techniques to increase the...
190

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A Strategy for Sensitive, Large Scale Quantitative Metabolomics
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Optimal Sample Preparation Workflow for Quantitative Mass Spectrometry-Based Studies in the Low Range.

Sara Kassem1,2, Paula Díez3,4,5

  • 1Department of Immunology, Leiden University Medical Center (LUMC), Leiden, The Netherlands.

Methods in Molecular Biology (Clifton, N.J.)
|December 23, 2024
PubMed
Summary

This study presents a standardized mass spectrometry (MS) workflow for analyzing limited cell samples. The protocol simplifies sample preparation for non-proteomics specialists, enabling TMT-based quantitative analysis.

Keywords:
Cell lysisCell storageMass spectrometryProtein quantificationProteomics sample preparationQuantity-limited sampleSP3 technologyTandem mass tag

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Molecular Biology

Background:

  • Mass spectrometry (MS) allows high-throughput protein characterization.
  • Advances enable deep proteomic analysis of low cell numbers, including single cells.
  • Standardized protocols are lacking for non-proteomics specialists.

Purpose of the Study:

  • To detail a reproducible and easy-to-implement workflow for cell-limited sample preparation.
  • To facilitate TMT-based relative quantitative MS analysis for a broader audience.
  • To address the need for standardized protocols in low-input proteomics.

Main Methods:

  • Cell storage and handling protocols.
  • Peptide labeling techniques.
  • TMT-based relative quantitative mass spectrometry analysis.

Main Results:

  • A standardized, easy-to-implement workflow for cell-limited sample preparation.
  • Successful TMT-based relative quantitative MS analysis from low cell numbers.
  • Reproducible results from the described protocol.

Conclusions:

  • The developed workflow simplifies low-input sample preparation for MS.
  • This protocol empowers non-proteomics specialists to perform quantitative proteomic analysis.
  • Standardization enhances the accessibility and reliability of single-cell and low-cell-number proteomics.