cOmplete™, EDTA-free Protease Inhibitor Cocktail
ROCHE/04693132001 - Tablets provided in EASYpacks
Synonym: protease inhibitor
Product Type: Chemical
cOmplete, EDTA-free
Figure 1: His-tagged hAkt1 protein purified and analyzed using SDS-PAGE followed by Coomassie Blue staining. hAkt1 is a challenging protein for expression at the 2-liter scale, because of low expression at the 6-well plate scale compared to another protein kinase. Therefore, protection using cOmplete EDTA-free Protease Inhibitor Cocktail Tablets was necessary to prevent degradation of the isolated protein. A total of 10 mg protein was purified from the 2-liter culture after transient transfection using the FuGENE HD Transfection Reagent, which is comparable to that reported for this protein in baculoviral systems (Lanes: 1: total lysates; 2: soluble; 3: flow through; 4-7: wash; 8-16: imidazole eluates of hAkt1).
Inhibition of different proteases by cOmplete Protease Inhibitor Tablets.
| absorption | 0.08 at 280 nm |
| form | tablet |
| manufacturer/tradename | Roche |
| packaging | pkg of 20 tablets (individually packed in foil blister packs) |
| solubility | aqueous buffer: soluble |
| water: soluble | |
| storage temp. | 2-8°C |
| 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 |
| Application: | 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 solution Working solution: Handling Carefully push the tablet through the foil packaging using the base of your thumb (not fingernail) to prevent the breakage of tablets. Preparation of Working Solution One 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) |
| Symbol |  GHS05 |
| Signal word | Danger |
| Hazard statements | H314 |
| Precautionary statements | P260 - P280 - P301 + P330 + P331 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 + P310 |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 2 |
| Flash Point(F) | does not flash |
| Flash Point(C) | does not flash |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352200 |