Protein Sequencing Sample Requirements

Proteomic sequencing is a technique for analyzing protein expression, modification, and interaction, which usually relies on mass spectrometry. To carry out effective proteomic analysis, the preparation of the sample must meet certain requirements. Here are some common proteomics sequencing sample requirements.

 

Purity of the Sample

The purity of the protein in the sample should be as high as possible to reduce interference from non-target proteins. It may be necessary to remove cellular debris, lipids, salts, and other small molecule impurities through centrifugation, filtration, precipitation, washing, and electrophoresis.

 

Quantity of the Sample

Usually, a sufficient amount of protein is required to ensure that enough peptides can be detected and identified. The specific amount depends on the sensitivity of the mass spectrometry equipment and the required depth of analysis, which may range from micrograms to milligrams.

 

Buffering of the Sample

The sample should be in a buffer solution suitable for mass spectrometry analysis. Ingredients of incompatible buffers, such as certain detergents, salts, or additives, may interfere with mass spectrometry analysis or damage the mass spectrometer.

 

Protein Modification

If certain modifications of the protein (such as phosphorylation, glycosylation, etc.) are the focus of the analysis, extraction and processing methods that protect these modifications should be used.

 

Stability

Protein samples should remain stable before analysis to avoid degradation or denaturation. This may require the sample to be processed and stored at low temperatures, and the use of protease inhibitors.

 

Sample Submission Information

Providing detailed sample information, including the type of sample, processing method, expected proteins to be detected, or the research objective, helps the laboratory better understand and meet the analysis needs.

 

Packaging and Transportation

The sample should be properly packaged and transported on ice packs or dry ice to avoid degradation or denaturation during transportation.

 

To ensure that the sample meets the requirements, the laboratory must follow strict standard operating procedures (SOPs) during the collection, processing, and transportation of the sample. In addition, it is best to communicate with the sequencing service provider before submission to confirm the specific sample requirements to avoid affecting the experimental results.