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luckyliu881011(金币+50, 翻译EPI+1):太感谢了了,先给了金币再看... 2010-10-17 07:58:05
The basic strategy is as following: in the quantitative amino acid sequences by selecting the size of a medium molecular weight (typically 0.9 ~ 2 kd), mass spectrometry peptide strong signal response as a template, using chemical synthesis or cell culture methods synthesized with a certain isotopes (eg 13C, 15N, 2H) peptide labeled the same (only with different molecular weight) as the internal standard, detection of peptides by mass spectrometry signal intensity ratio (peak height or peak area ratio) of absolute quantification of the corresponding proteins. Application of the standard method provided that: 1) The absolute quantification of protein digestion to fully; 2) It was chosen as the internal standard template peptide between peptides and proteins there is a certain quantitative relationship.
This method's strategy is: to be the determination of protein in acid conditions, full hydrolysis, all the formation of amino acid mixture, then add a known amount of isotope labeled amino acid as the internal standard by LC-MS/MS mass spectrometry one pair amino acids in order to achieve mass peak intensity of amino acid concentrations were measured, and then based on this amino acid in the protein content, calculate the amount of protein. |
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