For life science research only. Not for use in
diagnostic procedures. FOR IN VITRO USE ONLY.
c p mplete, 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
N Do not eat Store at +2 to +8°C
1.What this Product Does
1.1Properties
c p mplete, Mini, EDTA-free tablets inhibit a broa
d spectrum of serin
e and cysteine proteases. In contrast to other c p mplete tablets they do not contain EDTA leaving the stability and the function o
f metal dependant proteins uneffected. The affinity purification of Poly-His tagged fusionproteins via IMAC (immobilized metal affinity 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 p mplete, Mini, EDTA-free con-tains both irreversible and reversible protease inhibitors. Metallo and aspartic proteases are not inhibited.
1.2Contents
30 individually packed c p mplete Protease Inhibitor Cocktail Tablets in foil blisters. Each tablet is sufficient for a volume of 10 ml solution. 1.3Stability
•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 Ϫ15 to Ϫ25° C.
1.4Application
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
2.1Handling
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.
N Do not eat the tablets.
2.2Preparation of Working Solutions
One tablet c p mplete, 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 c p mplete, EDTA-free (1 tablet for 50 ml extraction solution, see "Ordering Information"). The composition of the c p m-plete, EDTA free tablet is identical to the c p mplete, Mini, EDTA-free tablet, therefore comparable results are achieved with both product types.2.3Stock solution (7 × conc.)
Dissolve one tablet c p mplete, Mini, EDTA-free in 1.5 ml redist. H2O or in 100 mM phosphate buffer, pH 7.0.
3.Results
c p mplete, Mini, EDTA-free tablets are applie
d to stabiliztablet5
e those extracts where the stability or activity o
f metal containin
g proteins should not be effected. Since EDTA interferes wit
h IMAC (immobilized metal affinity chromatography) c p mplete, Mini, EDTA-free is prefer-encially used in the isolation process of Poly-His tagged fusion pro-teins. c p mplete, 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 c p mplete, Mini, EDTA-free are shown in table 1.
Table 1:Inhibition of different proteases by c p mplete Protease Inhi-bitor Tablets.
One c p mplete, EDTA-free tablet was added per 10 ml incubation solution. Pro-teolytic activity was det
ermined 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 c p mplete, EDTA-free tablets to ensure aspartic (acid) protease inhibition. All experiments were performed at room temperature.
* available from Roche Applied Science
Protease
resp.
protease
mixture
Enzyme
concen-
tration
(mg/ml)
pH-
value
% inhibition
after
immediate
addition
to the
protease
% inhibition
after 60 min
incubation
(protease +
c p mplete)
at +15 to
+25° C Pancreas-
extract
0.027.884%94%
Thermolysin
(Metallopro-
tease)
0.00057.87%10%
Chymotrypsin0.002
0.02
7.8
7.8
90%
60%
100%
98%
Trypsin0.02
0.002
7.8
7.8
70%
81%
74%
86%
Papain0.337.874%87%
1205.0472*******➁
Roche Applied Science
Roche Diagnostics GmbH Roche Applied Science 68298 Mannheim Germany
4.Additional Information on this Product
4.1Absorption
A solution of 1 tablet c p mplete, Mini, EDTA-free dissolved in 10 ml H 2O has an absorption of 0.08 at 280 nm.
4.2Quality Control
The inhibitory activity of each lot is tested with a concentrated pan-creas extract. Thereby, the proteolytic activitiy is typically inhibited by 94% after one hour (detection with universal protease substrate,casein, resorufin-labeled*).
4.3Reference
1North, M.J. (1969) in: Proteolytic Enzymes - A Practical
Approach (Beynon, P.J. & Bond, J.S. eds.), IRL press Oxford, pp. 117-119.
5.Ordering Information
For addional information, please visit
c p mplete  Protease Inhibitor Cocktail Tablets in glass vials:
Kits and Sets:
Substrate:
Product Pack size
Cat. No.c p mplete 20 tablets in foil blisters (for 50 ml each)04 693 116 001c p mplete, Mini 30 tablets in foil blisters (for 10 ml each)04 693 124 001c p mplete, EDTA-free
20 tablets in foil blisters (for 50 ml each)
04 693 132 001c p mplete, Mini, EDTA- free 30 tablets in foil blisters (for 10 ml each)
04 693 159 001
Product Pack size
Cat. No.c p mplete
20 tablets in a glass vial
(for 50 ml each)
3 × 20 tablets in a glass vial (for 50 ml each)11 697 498 00111 836 145 001c p mplete, Mini 25 tablets in a glass vial (for 10 ml each)11 836 153 001c p mplete, EDTA-free
20 tablets in a glass vial (for 50 ml each)
11 873 580 001c p mplete, Mini, EDTA- free 25 tablets in a glass vial (for 10 ml each)
11 836 170 001
Product Pack size
Cat. No.
Pefabloc SC PLUS
Set I: contains 100 mg Pefabloc SC and 5 ml PSC protector solution Set II: contains 1g Pefabloc SC and 2 × 25 ml PSC protector solution
11 873 601 001
11 873 628 001Protease Inhibitor Set
Small quantities of 10 most commonly used protease inhibi-tors
11 206 893 001Product
Pack size Cat. No.Universal Protease Substrate (Casein, resorufin-labeled)
15 mg 40 mg
11 080 733 00111 734 334 001
Individual Protease Inhibitors:
This combination comprises only the most important products related to the product described.
Please refer to our latest catalog for our current product range or contact your local Roche Applied Science representative directly.
C p MPLETE is a Trademark of Roche
PEFABLOC is a Trademark of Pentapharm AG, Basel, Switzerland.
Contact and Support
To ask questions, solve problems, suggest enhancements or report new applications, please visit our  Online Technical Support Site  at:
To call, write, fax, or email us, visit the Roche Applied Science home page, he-applied-science, and select your home country. Country- specific contact information will be displayed. Utilize the Product Search function to get access to Pack Inserts and Material Safety Data Sheets.
Product Pack size Cat. No.Aprotinin
10 mg 50 mg 100 mg 10 236 624 00110 981 532 00111 583 794 001Bestatin 10 mg 50 mg 10 874 515 00111 359 070 001Calpain Inhibitor I 25 mg 11 086 090 001Calpain Inhibitor II 25 mg 11 086 103 001Chymostatin 10 mg 11 004 638 001E-64
5 mg 10 mg 25 mg 11 585 673 00110 874 523 00111 585 681 001Leupeptin
5 mg 25 mg 50 mg 100 mg 11 017 101 00111 017 128 00111 034 62
6 00111 529 048 001α2-Macroglobulin 25 IU 10 602 442 001Pefabloc SC
100 mg 500 mg 1 g 11 429 868 00111 585 916 00111 429 876 001Pepstatin
2 mg 10 mg 50 mg 10 25
3 286 00111 359 053 00111 52
4 488 001PMSF
1 g 10 g 25 g 10 236 608 00110 837 091 00111 359 061 001TLCK – HCl 100 mg 250 mg 10 874 485 00110 874 493 001Trypsin Inhibitor (chicken, egg white)  1 g 10 109 878 001Trypsin Inhibitor (soybean)
50 mg 500 mg
10 109 886 00110 109 894 001

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