Description
General description
Recombinant proteins may be partially degraded during expression. Soluble degradation products are co-purified and cause a non-homogeneous protein preparation with protein fragments of different lengths. However, metalloproteases are prevalent in (for example) bacteria and some techniques require that EDTA be omitted.
cOmplete ULTRA Tablets, Mini, EDTA-free contain the most powerful protease inhibitor cocktail for the optimal protection of your proteins against protease degradation. They are supplied as tablets in an EASYpack blister pack. EASYpacks provide the most convenient form of delivery - single tablets are easily pushed through the blister foil directly into the buffer. One cOmplete ULTRA Tablet, Mini, EDTA-free is sufficient for a volume of 10 ml extraction solution.
cOmplete ULTRA Tablets, Mini, EDTA-free are identical to cOmplete ULTRA Tablets, Mini, the only difference being that no EDTA or other chelating agents (e.g. EGTA) are included.
Application
During cell lysis and purification, target proteins are at risk of being cleaved by proteases, which are naturally present in cells. To ensure that full-length proteins are obtained, cOmplete™ ULTRA Tablets are used for the inhibition of an extended range of proteases. This new formulation shows highest inhibition ratios due to its broadened range of inhibitors compared to standard cOmplete Tablets.
This new protease inhibitor cocktail now inhibits aspartic proteases, as well as serine and cysteine proteases. cOmplete ULTRA Tablets were developed for maximum efficiency on different cell types. Best performance is guaranteed in lysates of bacteria, yeast, insect cells, and a wide range of higher cell lines.
This tablet does not contain EDTA; therefore, metalloproteases are not inhibited.
For the purification of histidine-tagged proteins using Immobilized Metal Ion Affinity Chromatography (IMAC), this EDTA-free tablet is recommended for applications using non-EDTA compatible IMAC resins. Alternatively, an EDTA-containing cOmplete ULTRA, Mini tablet may be used in combination with Roche ′s new EDTA-compatible chromatography material, cOmplete His-Tag Purification Resin.
cOmplete™ ULTRA Tablets, Mini, EDTA-free, EASYpack Protease Inhibitor Cocktail has been used as a component of
homogenization buffer for muscle samples for immunoblotting sample preparation
radioimmunoprecipitation assay buffer (RIPA buffer) for protein extraction from brain samples for western blotting
lysis buffer for the human embryonic kidney 293 (HEK293) lysate preparation for western blotting
Biochem/physiol Actions
Inhibits serine proteases, cysteine proteases, and aspartic proteases. Does not inhibit metalloproteases.
Features and Benefits
Protect your precious proteins.
Choose cOmplete ULTRA for even more efficient protease inhibition in eukaryotic, yeast, and bacterial cell lysates.
Why settle for less.
Benefit from 7 powerful protease inhibitors – more than in other EDTA-free inhibitor tablets.
Defend against a broad range of proteases.
Reliably inhibit serine, cysteine, and aspartic proteases.
Convenient and safe tablet format.
Eliminate weighing small amounts of individual inhibitors which might be hazardous. Simply drop a tablet from the blister into your buffers.
Physical form
Tablets in foil blister packs. Each tablet is sufficient for a volume of 10 ml extraction solution.
Preparation Note
Working concentration: 1 tablet per 10 ml extraction solution
Working solution: Handling
Carefully push the tablet through the foil packaging using the base of your thumb (not your fingernail) to prevent the breakage of tablets.
Preparation of Working Solution
One tablet is sufficient for the inhibition of proteolytic activity in 10 ml extraction solution. If very high proteolytic activity is present, 2-3 tablets should be used for 10 ml extraction buffer. The tablets can be added directly to the extraction medium.
Stock Solution (2x conc.)
Alternatively to the direct use with lysates, a 2x stock solution can be prepared, with the following procedure:
• Add one cOmplete ULTRA Tablet, Mini, EDTA-free to 5 ml double-distilled water in a PE or PP tube.
• Vortex 3 min or until the tablet is completely dissolved.
• The stock solution is ready to use even if there are very fine particles still visible in solution. These particles do not interfere with the inhibition performance of the tablets.
• This stock solution is stable for 4 weeks, stored at 2 to 8 °C, and for at least 4 weeks at -15 to -25 °C as well.
• During storage of the 2x stock solution, the particles can sediment. Briefly vortex the stock solution prior to use.
Storage conditions (working solution): The stock solution in water is stable for 4 weeks, stored at 2 to 8 °C, and for at least 4 weeks at -15 to -25 °C as well.
Soluble in aqueous buffers, or add directly to extraction media. For the stock solution, use water only.
Other Notes
For life science research only. Not for use in diagnostic procedures.
Proprietary mixture of several protease inhibitors with broad inhibitory specificity.
Legal Information
cOmplete is a trademark of Roche
