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蛋白酶抑制剂混合物Cocktail(Roch公司产品)

发布时间 2018-01-19
 

cpmplete, Mini, EDTA-free     
Protease Inhibitor Cocktail Tablets provided in  EASYpack  For the complete inhibition of serine and cysteine proteases during extractions from animal and plant tissues or cells, yeast and bacteria  
Cat. No. 04 693 159 001   30 tablets Version June 2005  Store at +2 to +8°C
1. What this Product Does
1.1 Properties
cpmplete, Mini, EDTA-free tablets inhibit a broad spectrum of serine and cysteine proteases. In contrast to other cpmplete tablets they do not contain EDTA leaving the stability and the function of metal dependant proteins uneffected. The affinity purification of Poly-His tagged fusionproteins via IMAC (immobilized metal ffinity chroma-tography) 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 therefore very well suited for the protection of proteins isolated from animal tis-sues, plants, yeast and bacteria. c pmplete, Mini, DTA-free  con-tains both irreversible and reversible protease inhibitors. Metallo and aspartic proteases are not inhibited. The protease inhibitors of cpmplete, Mini, EDTA-free do not form irreversible complexes with thiol (SH) groups of proteins. 
1.2 Contents
30 individually packed c pmplete Protease Inhibitor Cocktail Tablets in foil blisters. Each tablet is sufficient for a volume of 10 ml solution. 
1.3 Stability
? The tablets are stable at +2 to +8°C, stored dry, until expiration date.
? The stock solution is stable for 1–2 weeks, stored at +2 to +8°C, or  at least for 12 weeks at  -15 to -25° C.
1.4 Application
Used for the inhibition of serine and cysteine proteases in bacterial, yeast, plant and animal cell extracts. For the inhibition of proteolytic activity in smaller volumes, in which EDTA may interfere with protein stability or subsequent assays or purification methods (e.g.,  Immobilized Metal Chelate Affinity Chro-matography [IMAC]).
2. How to Use this Product
2.1 Handling
Carefully push the tablet through the foil packaging using the base of  your thumb (and not yout finger nail) to prevent the breakage of tab-lets. 
2.2 Preparation of Working Solutions
One tablet c pmplete, Mini, EDTA-free is sufficient for the inhibition of the proteolytic activity in 10 ml extraction solution. If very high pro-teolytic activity is present, one tablet should be used for 7 ml extraction buffer. The tablets can be added directly to the extraction medium. Alternatively a stock solution (7 × conc.) can be prepared. 
If it is necessary to inhibit proteolytic activity in a larger volume we rec-ommend to use cpmplete, EDTA-free (1 tablet for 50 ml extraction solution, see "Ordering Information"). The composition of the cpm-plete, EDTA free tablet is identical to the c pmplete, Mini, EDTA-free tablet, therefore comparable results are achieved with both product type
2.3 Stock solution (50 × conc.)
Dissolve one tablet cpmplete, Mini, EDTA-free in 0.2 ml redist. H2O or in 100 mM phosphate buffer, pH 7.0.
3. Results
cpmplete, Mini, EDTA-free tablets are applied to stabilize those extracts where the stability or activity of metal containing proteins should not be effected. Since EDTA interferes with IMAC (immobilized metal affinity chromatography) cpmplete, Mini, EDTA-free is prefer-encially used in the isolation process of Poly-His tagged fusion pro-teins. c pmplete, Mini, EDTA-free tablets inhibit efficiently serine and cysteine proteases in a broad range but not metalloproteases. Occasionally, aspartic proteases (“acid proteases“) can interfere upon isolations from animal tissues or bacterial extracts (1). These pro-teases, however, exhibit pronounced activities only at low pH values. If extraction or single isolation steps have to be performed at this pH range the addition of pepstatin* is recommended to inhibit aspartic protease activity as a precaution. Typical values for the inhibition of different proteases and protease mixtures by cpmplete, Mini, EDTA-free are shown in .
 
One cpmplete, EDTA-free tablet was added per 10 ml incubation solution. Pro-teolytic activity was determined with the Roche Applied Science Universal Pro-tease Substrate (casein, resorufin-labeled*). When extractions or single-step isolations are necessary in the acid pH range, simply include pepstatin* along with cpmplete, EDTA-free tablets to ensure aspartic (acid) protease inhibition. All experiments were performed at room temperature.  
* available from Roche Applied S
 
Origin:     Roche  Biotechnology,  Inc of  USA
Distributor: Pioneer Biotechnolgy,Inc 
Contact:  Tel 029-88237772  or 13484545009  Fax 029-88237772
  E-mail : xfyangbio@163.com      Internet: www.xfbio.com

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