Not available outside of the UK & Ireland.

Application

Use Chymotrypsin Sequencing Grade for the hydrolysis of proteins by chymotrypsin alone or in combination with other proteases. It is suitable for peptide mapping, fingerprinting, and sequence analysis.

General description

Chymotrypsin Sequencing Grade is isolated as a specific protease in pure form from bovine pancreas.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Preparation Note

Working concentration: The recommended amount of enzyme is 1/200 to 1/20 of the quantity of protein by weight.Storage conditions (working solution): 2 to 8 °CA solution in 1 mM HCl can be stored for up to one week at 2 to 8 °C.Inhibitors: Aprotinin, DFP, PMSF, phenothiazine-N-carbonyl chloride, TPCK, ZPCK, α2-macroglobulin, α1-antitrypsin, soybean trypsin inhibitor, and chymostatin. No inhibition by APMSF.

Quality

Purity: Free of impurities that may interfere with the specific cleavage or separation of peptides in reversed-phase HPLC.

Reconstitution

Dissolve lyophilized chymotrypsin sequencing grade in 1mM HCl. The proteins to be sequenced are dissolved in digestion buffer (100mM Tris-HCl, 10mM CaCl₂, pH 7.8).

Specificity

Serine endopeptidase that specifically hydrolyzes peptide bonds at the C-terminial of Tyr, Phe, and Trp. Leu, Met, Ala, Asp, and Glu are cleaved at a lower rate. Acts also upon amides and esters of susceptible amino acids. The specificity of Chymotrypsin Sequencing Grade is tested with melittin as a substrate.