Trypsin (Porcine)

 

Trypsin (Porcine)

Trypsin is a serine endopeptidase which specifically cleaves peptide bonds on the carboxy side of arginine, lysine and s-aminoethyl cysteine residues.

Characteristics:

Trypsin is a serine endopeptidase which specifically cleaves peptide bonds on the carboxy side of arginine, lysine and s-aminoethyl cysteine residues. Typically there is little or no cleavage at arginyl-proline or lysyl-proline bonds. Princeton Separations sequencing grade trypsin, TPCK treated, is subjected to extensive purification to remove contaminating proteases and tryptic autolysis by-products which could affect the specificity of the digestion process. Subsequently the enzyme is modified chemically to minimize autolysis as well as increase the stability.

Application:

Trypsin, Sequencing Grade, modified is supplied lyophilized in 25 µg vials. The enzyme is typically reconstituted to a concentration of 1 µg/µL with deionized H2O or buffer of choice (25 µL per vial). The modified enzyme retains 70-90% of ts activity after 6 hours incubation at 30OC in reaction buffer and 65 - 80% of its activity after 24 hours under the same conditions. To use, the enzyme is added to the sample protein to a ratio of 1/100 to 1/20 (enzyme to protein, by weight) in a standard digestion buffer such as 50mM Tris-HCI pH 8.0. Ammonium carbonate buffers are slightly inhibitory and are not recommended. The incubation is allowed to proceed at 25 - 30OC for 1-10 hours, but can extend to 24 hours in some applications. The Princeton Separations enzyme is stable for at least 24 hours at 30OC and the autodigestion products are minimal. However, it is always preferable to use the shortest incubation time possible, since cleavage, particularly on the carboxy side of hydrophobic residues, has been found to occur following prolonged incubations (>8 hours). Optimal incubation time can be reached by adjustment of the enzyme to sample ratio.

Quality Control:

Two assays are used for Quality Control: an amidase assay and a protease (digestion) assay using casein as substrate. The activity against casein is routinely compared with that of Trypsin and a Trypsin Activity Equivalence is calculated. Enzyme specificity is checked using an array of synthetic peptides.

Price and ordering information:

Product must be shipped 2 day air

Price: $0.00
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Procedure Guide for Trypsin (Porcine)46.86 KB

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