Immobilized Chymotrypsin (CTRC) Active Protein

Bovine Pancreas

Resin; 0.02% NaN3 50% Glycerol

>95% by SDS-PAGE analysis

Ships on blue ice. Store at 4°C.

25,000

ModBase 3D Structure for Q7M3E1

Protein activity can be an ambiguous term. Please refer to the data sheet or inquire with Technical Support to determine the definition of activity for this product.

Sodium Azide

180 TAME U/ml (1 Unit is equal to 1 umol of TAME (p-toluenesulfonyl-L-arginine methyl ester) hydrolyzed per minute at pH 8.2, 25°C)

1. Make a digestion buffer consisting of 0.08M Tris-HCl, 0.1M Calcium Chloride, pH 7.8 (or other suitable buffer such as 0.1M Ammonium Bicarbonate, 0.01M Calcium Chloride, pH 8.0).
2. Dissolve 1 mg of your protein sample in 0.2ml digestion buffer.
3. Wash 0.2-0.3ml of immobilized chymotrypsin with 3x0.5ml of digestion buffer. Separate the gel from the buffer after each wash by centrifugation or by column. Discard buffer after each wash.
4. Resuspend the washed immobilized chymotrypsin gel in ~0.2ml of digestion buffer then add the resin to your protein sample.
5. Incubate mixture in a shaking water bath for 2-18 hours at 37°C with rapid agitation.
6. Separate the immobilized chymotrypsin gel from the digestion mixture as noted in step 3. Retain supernatant as your chymotrypsin-digested protein sample.

Note: Optimization of immobilized chymotrypsin protocol is required for specific applications. Recommended reaction conditions are pH 7.5 to 9.0 at 37°C. The reaction rate will be increased by increasing the enzyme to protein substrate ratio and incubation temperature. For example, a typical digestion of 4 hours to overnight can be achieved using a ratio of 1:25 enzyme to protein substrate, while it is recommended to use 1:10 enzyme to protein substrate for accelerated chymotrypsin digestion (30-60 minutes).