General Protocol for Amino Acid Sequence Determination of Proteins

GENERAL PROTOCOL FOR AMINO ACID SEQUENCE DETERMINATION OF PROTEINS 

Homogenization of the biological sample (target tissue, biofluid, cell culture, or microorganism). 

Separation of the target protein(s) using one- or two-dimensional sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) or HPLC techniques. 

Reduction-alkylation of proteins. The molecular mass measurement of the target protein before and after this step provides information on the number of cysteine residues and disulfide bonds. 

Site-specific cleavage of reduced and alkylated proteins into smaller, more manageable peptides by chemical degradation or proteolysis. 

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