Application

cOmplete^™, EDTA-free Protease Inhibitor Cocktail has been used as a component in lysis buffer for the lysis of transfected human embryonic kidney (HEK293T) cells and in radioimmunoprecipitation assay (RIPA) lysis buffer for the homogenization of zebrafish embryo tissue protein extraction reagent (T-PER) for the homogenization of hippocampus tissue sodium dodecyl sulphate (SDS) lysis buffer for the lysis of hepatocytes

cOmplete^™, EDTA-free, efficiently inhibits serine and cysteine proteases in bacterial, yeast, plant, and animal cell extracts. Metallo- and aspartic proteases are not inhibited.cOmplete, EDTA-free Tablets, are used for the inhibition of proteolytic activity in large volumes (up to 50ml) in which EDTA may interfere with protein stability (e.g., metal-containing proteins) or subsequent assays. Since EDTA interferes with IMAC (Immobilized Metal Affinity Chromatography), cOmplete, EDTA-free is preferentially used in the isolation process of Poly-His-tagged fusion proteins.If it is necessary to inhibit proteolytic activity in a smaller volume (up to 10ml), we recommend to use cOmplete, Mini, EDTA-free.One cOmplete, EDTA-free tablet was added per 50 ml incubation solution. Proteolytic activity was determined with the Roche Universal Protease Substrate (casein, resorufin-labeled). When extractions or single-step isolations are necessary in the acid pH range, simply include pepstatin along with cOmplete, EDTA-free tablets to ensure aspartic (acid) protease inhibition. All experiments were performed at +15 to +25°C.

Components

Proprietary mixture of several protease inhibitors with broad inhibitory specificity.

Features and Benefits

Use cOmplete Protease Inhibitor Tablets to protect your proteins from a wide range of proteases. In just minutes, uninhibited proteolytic activity can degrade the protein you have spent days isolating. Each of these convenient water-soluble tablets contains a blend of protease inhibitors that inhibits proteolytic activity from most cell types, including animals, plants, yeast, and bacteria. No weighing or measuring is necessary, ensuring consistent results. Convenience: Simply push the quick-dissolving tablet through the foil packaging of the EASYpack into your buffer. Complete protection: Instantly protect your proteins against a broad range of proteases. Flexible: Protect proteins in extracts from almost any tissue or cell, including animals, plants, yeast, bacteria, or fungi. Safe: Choose non-toxic inhibitors that pose no risk to you or those around you. Suitable for IMAC purification: Purify poly-histidine tagged proteins with cOmplete His-Tag Purification Resin.

General description

cOmplete, EDTA-free Protease Inhibitor Tablets, inhibit a broad spectrum of serine and cysteine proteases. In contrast to other cOmplete tablets they do not contain EDTA - no other chelating agents, e.g. EGTA as well - leaving the stability and the function of metal-dependant proteins unaffected. The affinity purification of Poly-His-tagged fusion proteins using IMAC (Immobilized Metal Affinity Chromatography) is also facilitated (no dialysis necessary).Due to the optimized composition of the tablets, they show excellent inhibition of serine and cysteine proteases, and are well suited for the protection of proteins isolated from animal tissues, plants, yeast, and bacteria. cOmplete, EDTA-free tablets, contains both irreversible and reversible protease inhibitors. Metallo- and aspartic proteases are not inhibited.cOmplete EDTA-free Tablets are identical to cOmplete Tablets, the only difference being that no EDTA or other chelating (e.g. EGTA) agents are included.

Legal Information

cOmplete is a trademark of Roche

Other Notes

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

Packaging

20 individually packed cOmplete Protease Inhibitor Cocktail Tablets in foil blisters.

Physical form

Tablets in foil blister packs

Preparation Note

Working concentration: 1 tablet per 50 ml extraction solutionWorking solution: HandlingCarefully push the tablet through the foil packaging using the base of your thumb (not fingernail) to prevent the breakage of tablets.Preparation of Working SolutionOne cOmplete EDTA-free tablet is sufficient for the inhibition of the proteolytic activity in 50 ml extraction solution. When very high proteolytic activity is present, one tablet should be used for 25 ml extraction buffer. Tablets can be added directly to the extraction medium. Alternatively, a stock solution (25 X conc.) can be prepared.Preparation of Stock solution (25 X conc.)Dissolve one cOmplete EDTA-free tablet in 2 ml dist. water or in 2 ml 100 mM phosphate buffer, pH 7.0.Storage conditions (working solution): The stock solution can be stored at 2 to 8 °C for 1 to 2 weeks, or at least 12 weeks at -15 to -25 °C.

Quality

Performance-tested with pancreas extract.

Reconstitution

Soluble in aqueous buffers, or add directly to extraction media. Alternatively, prepare 25x stock solutions in 2 ml water or 100 mM phosphate buffer, pH 7.0. Stock solution is stable for 1-2 weeks at 2 to 8 °C or at least 12 weeks at -15 to -25 °C; an be used in thiol-containing solutions at 15 to 25 °C.

Specificity

Mixture of several protease inhibitors. For the inhibition of serine, cysteine, but not metalloproteases.

Storage and Stability

Store at 2 to 8 °C. (storage at -15 to -25 °C is also possible)