Cross-Linking Mass Spectrometry Sample Preparation
Cross-linking mass spectrometry is an effective tool for studying protein structure and function. During the experimental process, sample preparation is a critical step. The main steps in cross-linking mass spectrometry sample preparation are protein extraction, protein purification, protein cross-linking, and preparation of cross-linked protein for mass spectrometry analysis.
1.Protein Extraction
First, the target protein is extracted from cells or tissues using protein extraction methods. The steps are as follows:
• Use a protein extraction buffer to lyse the cells or tissues.
• Centrifuge to remove cell or tissue debris.
• Transfer the supernatant to a new centrifuge tube and measure the protein concentration using a protein assay kit.
2. Protein Purification
Next, the extracted protein needs to be purified. The steps are as follows:
• Purify the protein using a protein purification kit.
• Check the quality and purity of the purified protein using SDS-PAGE electrophoresis.
3. Protein Cross-linking
Next, cross-link the protein with a cross-linking agent. The steps are as follows:
• Add an appropriate amount of cross-linking agent to the protein solution and allow it to react.
• After the reaction, neutralize the reaction mixture with a buffer.
4. Preparation of Cross-linked Protein for Mass Spectrometry
Finally, prepare the cross-linked protein for mass spectrometry analysis. The steps are as follows:
• Digest the cross-linked protein into peptides using a kit.
• Conduct mass spectrometry analysis using LC-MS/MS.
It should be noted that the above steps may need to be adjusted depending on the protein and cross-linking agent used. Additionally, the quality of sample preparation directly affects the results of mass spectrometry analysis, so it is crucial to perform these operations carefully.
BiotechPack, A Biopharmaceutical Characterization and Multi-Omics Mass Spectrometry (MS) Services Provider
Related Services:
Cross-linking Protein Interaction Analysis
Protein Interaction Mass Spectrometry Analysis
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