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广州市左克生物科技发展有限公司
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文献和实验Partial proteolysis with Staph. aureus V8 protease
mM Tris-HCl, pH 6.8, and 1mM EDTA. This is in Tilo's box 3, in the east Forma, #1. Generally, if you aren't sure how much enzyme to use, try 10, 50 and 100 nanograms of V8 protease. If V8 doesn't work for your protein, try papain
Two dimensional peptide mapping
so that you have at least 500 cpm. Transfer the protein to nitrocellulose, Immobilon or Nylon according to Kamps's Western Protocol . Tryptic digestion can be done with either NC, Nylon, or Immobilon. Nitrocellulose seems to give more reproducible yields of peptides.
Byonic: Advanced Peptide and Protein Identification Software
package for peptide and protein identification by tandem mass spectrometry. This software, which has only recently become commercially available, facilitates a much wider range of search possibilities than previous search software such as SEQUEST
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