1. SILAC outlines SILAC, ie isotope labeling with amino acids in cell culture (SILAC). The basic principle is to use cultures containing light, medium and heavy isotopic essential amino acids in cell culture. The cell is cultured. The main stable isotope amino acids for SILAC are Lys and Arg. During the subculture, the isotope amino acids will be used by the cells to synthesize proteins. After subculture for 5-6 generations, all the proteins of the cells will be Isotopic labeling, equal-mixing of labeled proteins, SDS-PAGE separation and mass spectrometry, relative quantification by comparing the size of the three isotopic peptides in the first-order mass spectrum, and sequencing of the peptides by the second-order spectrogram Thereby identifying proteins.
Because SILAC labeling technology is an in vivo labeling technology that has little effect on cell function and high sensitivity, it is widely used in proteomics related fields, such as comparative proteomics, protein-protein interaction, protein and DNA. Interactions, proteins and RNA interactions and other fields.
2. Compared with in vitro labeling technology, SILAC is an in vivo marker with the following technical advantages: 1. High throughput, which can simultaneously identify and quantify hundreds to thousands of proteins;
2, quantitative accuracy, reduce experimental differences caused by different sample preparation;
3. Wide linear quantitative range;
4. The sensitivity is higher and the protein requirement is significantly reduced;
5, the mark uses the in vivo marking technology, closer to the true state of the sample.
Third, SIALC technical service process![]()
Fourth, SILAC technical service content 1, cell isotope labeling;
2. Sample preparation and quantification;
3. Separation by SDS-PAGE;
4. Liquid phase separation and mass spectrometry after trypsin digestion;
5. Database search; quantitative analysis of protein;
6. Relevant bioinformatics analysis;
7. Submit the experimental report.
V. Instructions for sending samples 1. Technical consultation: Before you start the experiment, the company will provide you with the most effective SILAC experimental design scheme. The company provides amino acids with Lys and Arg as L, M and H.
2. In order to ensure the smooth progress of the SILAC experiment, the sample delivery must meet the following requirements:
Basic requirements: SILAC experiments are only applicable to materials that can be cultured in SILAC medium. Please consult our company for details.
Transportation requirements: Liquid nitrogen or dry ice is stored and sent, and local customers in Guangzhou can pick up the samples.
Note: Samples should avoid all types of pollution and repeated freezing and thawing.
6. Experimental report content 1. Protein identification and SILAC quantitative analysis results;
2. The mass spectrum peak map corresponding to the protein specified by the customer;
3, SILAC experiments complete experimental procedures, the use of instruments, software retrieval parameters.
Seven, SIALC technical service case
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Eight, quality commitment 1, rigorous SILAC experimental design program to ensure reliable results of quantitative analysis;
2. Provide objective SILAC experimental analysis results.
Because SILAC labeling technology is an in vivo labeling technology that has little effect on cell function and high sensitivity, it is widely used in proteomics related fields, such as comparative proteomics, protein-protein interaction, protein and DNA. Interactions, proteins and RNA interactions and other fields.
2. Compared with in vitro labeling technology, SILAC is an in vivo marker with the following technical advantages: 1. High throughput, which can simultaneously identify and quantify hundreds to thousands of proteins;
2, quantitative accuracy, reduce experimental differences caused by different sample preparation;
3. Wide linear quantitative range;
4. The sensitivity is higher and the protein requirement is significantly reduced;
5, the mark uses the in vivo marking technology, closer to the true state of the sample.
Third, SIALC technical service process
Fourth, SILAC technical service content 1, cell isotope labeling;
2. Sample preparation and quantification;
3. Separation by SDS-PAGE;
4. Liquid phase separation and mass spectrometry after trypsin digestion;
5. Database search; quantitative analysis of protein;
6. Relevant bioinformatics analysis;
7. Submit the experimental report.
V. Instructions for sending samples 1. Technical consultation: Before you start the experiment, the company will provide you with the most effective SILAC experimental design scheme. The company provides amino acids with Lys and Arg as L, M and H.
2. In order to ensure the smooth progress of the SILAC experiment, the sample delivery must meet the following requirements:
Basic requirements: SILAC experiments are only applicable to materials that can be cultured in SILAC medium. Please consult our company for details.
Transportation requirements: Liquid nitrogen or dry ice is stored and sent, and local customers in Guangzhou can pick up the samples.
Note: Samples should avoid all types of pollution and repeated freezing and thawing.
6. Experimental report content 1. Protein identification and SILAC quantitative analysis results;
2. The mass spectrum peak map corresponding to the protein specified by the customer;
3, SILAC experiments complete experimental procedures, the use of instruments, software retrieval parameters.
Seven, SIALC technical service case
Eight, quality commitment 1, rigorous SILAC experimental design program to ensure reliable results of quantitative analysis;
2. Provide objective SILAC experimental analysis results.
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