cOmplete™, Mini, EDTA-free Protocol

Product No. 11836170001

Protocol

Complete, Mini, EDTA-free: How much to add to serum samples?

Dissolve one tablet in 10 mL aqueous buffer or water. If high proteolytic activity is present, dissolve one tablet in 7 mL buffer.

cOmplete Inhibition of Endoproteinase Asp-N

cOmplete inhibits the metalloprotease endoproteinase Asp-N.

cOmplete tablets for use in preparing subcellular fractions from intestine

This has not been tested, but should work.

Using cOmplete tablets in cell culture

An application could be to express/produce a protein which is very protease sensitive.

cOmplete Protease Inhibitor Cocktail can be used to inhibit proteases in the medium/supernatant of cell cultures. However, Roche has not systematically tested this application. cOmplete Tablets contain components which are membrane permeable, but not all are membrane permeable. In addition, the low molecular weight components which can enter cells have not been tested with regard to cytotoxicity.

Several customers have reported using cOmplete tablets successfully in cell culture at a dilution of one-third to one-fourth of the working concentration indicated in the package insert. To determine the suitability of cOmplete tablets for your specific application empirically test different concentrations.

Effect of cOmplete Inhibitor on nucleases such as DNase I and RNase A

Roche has not systematically tested the effect of cOmplete Protease Inhibitor Cocktail Tablets on DNase I and RNase A.

EDTA is a divalent chelator and can inactivate nucleases, such as Mg²⁺ required for DNase I activity. While low concentrations of EDTA are present in the reaction buffers of many restriction endonucleases, it may be best to use cOmplete EDTA-free Tablets.

Effect of cOmplete on phosphatases

cOmplete Protease Inhibitor Cocktail Tablets have not been systematically tested for inhibition of phosphatases by Roche.

EDTA is known to inhibit alkaline phosphatase, so as indicated above, it may be best to use cOmplete EDTA-free Protease Inhibitor Tablets.

Removing cOmplete from preparations using dialysis

Use a dialysis membrane with a cut-off of >10 kDa.