STEP 1 and 2 Chemical Method

STEP 1 and 2 Chemical Method

1. is done either by endopeptidases such as trypsin or pepsin .

2. peptides longer than c.50-70 aa can't be seq reliably by Edman Degredation.

3. long protein chains need to be broken up into small fragments

   • can then be seq individually

4. DIGESTION

   • or by chemical reagents such as cyanogen bromide
   • Different enzymes give different cleavage patterns, and the overlap between fragments can be used to construct an overall sequence
     • PURIFY PEPTIDES
       • REVERSE PHASE HIGH PERFORMANCE LIQUID CHROMATOGRAPHY