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For life science research only. 
Not for use in diagnostic procedures.
Protease Inhibitors Set
Cat. No. 11 206 893 001
1 set
y Version 17
Content version: August 2014
Product description
The set contains ten protease inhibitors. When used in varying combinations, a broad spectrum of proteases can be inhibited. For isolation and purification of
proteins (e.g. enzymes), specific mixtures of inhibitors can be used, for example a cocktail of Aprotinin, E-64 or Leupeptin, EDTA, Pefabloc® SC and Pepstatin.
Alternatively, our range of Complete Protease Inhibitor Cocktail Tablets provide a convenient and reliable solution for the inhibition of a broad spectrum of
serine, cysteine, metalloproteases as well as calpains. Due to the optimized composition of the tablets they show excellent inhibition, effects and are therefore
very well suited for the protection of proteins isolated from animal tissue, plants, yeast and bacteria. See below for ordering information.
Table 1: Composition of the set:
Suggested starting concentration ➀
Specificity of Inhibitors
Solubility/ Stability
Antipain-dihydrochloride (1)
3 mg
677.6
papain, trypsin
soluble in H2O (20 mg/ml),
methanol or DMSO ➁
50 ␮g/ml (74 ␮M)
Bestatin (1)
0.5 mg
308.4
amino peptidase, including aminopeptidase B., leucine aminopeptidase,
tripeptide aminopeptidase
soluble in 1 M HCI (20 mg/ml).
Methanol (5 mg/ml) or 0.15 M
NaCI (1 mg/ml). ➂
40 ␮g/ml (130 ␮M)
Chymostatin (3)
1 mg
E-64 (4)
3 mg
607.7
-, -, -, -chymotrypsin
357.4
cysteine proteases
soluble in glacial acetic acid
(20 mg/ml) or DMSO ➁
soluble in 1:1 H2O/ethanol 
[v/v] (20 mg/ml) ➁
Leupeptin (3)
0.5 mg
475.6
serine and cysteine proteases
such as plasmin, trypsin,
papain, cathepsin B
soluble in
H2O (1 mg/ml) ➁
Pepstatin (3)
0.5 mg
685 9
aspartate proteases like pepsin, renin,
cathepsin D, chymosin
soluble in
methanol (1.0 mg/ml) ➁
Phosphoramidon (3)
3 mg
543.5
soluble in
metallo endopeptidases, specifically
thermolysine, collagenase, metallo endo- H2O (20 mg/ml) ➁
proteinases.
Pefabloc® SC (6)
20 mg
239.5
serine proteases, e.g. trypsin,
chymotrypsin, plasmin, thrombin
EDTA-Na2 (5)
10 mg
372.24 metalloproteases
Inhibitor/Quantity
Aprotinin (2) 0.5 mg
➀
Mr
6512
serine proteases
6– 60 ␮g/ml (10–100 ␮M)
0.5– 10 ␮g/ml
(1.4–28 ␮M)
0.5– 5 ␮g/ml (1– 10 ␮M)
0.7 ␮g/ml (1 ␮M)
4– 330 ␮g/ml
(0.007– 0.6 mM)
soluble in
H2O (100 mg/ml) ➁
0.1– 1 mg/ml
(0.4– 4 mM)
soluble in
H2O to 0.5 M at pH 8-9.➁
0.2-0.5 mg/ml
(0.5-1.3 mM)
soluble in
H2O (10 mg/ml) ➂
(0.06– 2 ␮g/ml
(0.009- 0.3 ␮M
We recommend that these values be used only as a starting point. Final suitable concentrations must be determined empirically for each new system.
➁ Stock solutions are stable for 1 month stored at ⫺15 to ⫺25° C; storage in aliquots is recommended to avoid repeated thawing.
➂ Stock solutions are stable for 6 months stored at ⫺15 to ⫺25° C: storage in aliquots is recommended to avoid repeated thawing
Note: The effectiveness of protease inhibition can be verified fast and sensitively with the general substrate for proteases casein, resorufin labeled* (7)
For the different sources of biological material specifically adapted protease inhibitor cocktails have been found valuable for protein isolation (table 2):
Table 2: Suggestions for inhibitor cocktalls for protein isolation modified from (8) and (9)].
Tissue/Organism
Animal tissues
Plant tissues
Protozoa
Slime moulds
Yeast and fungi
Bacteria
Inhibitor
Pefabloc SC
EDTA
Leupeptin
Pepstatin
Aprotinin
Pefabloc SC
Chymostatin
EDTA
Pefabloc SC
Leupeptin
Pefabloc SC
Leupeptin
TLCK
Pefabloc SC
Phenanthroline
Leupeptin (9)
Pepstatin
Pefabloc SC
EDTA
Final concentration
1 mg/ml
0.5 mg/ml
10 ␮g/ml
10 ␮g/ml
1 ␮g/ml
1 mg/ml
20 ␮g/ml
0.5 mg/ml
1 mg/ml
10 ␮g/ml
1 mg/ml
10 ␮g/ml
0.04 mg/ml
1 mg/ml
1 mg/ml
3 ␮g/ml
15 ␮g/ml
1 mg/ml
0.5 mg/ml
Stock medium
water/buffer
water/buffer
water/buffer
methanol
water/buffer
water/buffer
DMSO
water/buffer
water/buffer
water/buffer
water/buffer
water/buffer
1 mMol HCI
water/buffer
ethanol
DMSA
methanol
water/buffer
water/buffer
Stock concentration
40 mg/ml
40 mg/ml
1 mg/ml
1 mg/ml
0.1 mg/ml
40 mg/ml
1 mg/ml
40 mg/ml
40 mg/ml
1 mg/ml
40 mg/ml
1 mg/ml
20 mg/ml
40 mg/ml
100 mg/ml
1 mg/ml
1 mg/ml
40 mg/ml
40 mg/ml
If cysteine proteases are thought to cause problems in animal, plant or protozoa tissues, E-64 should be checked as additive (final concentration: 10 ␮g/ml; stock solution: 1 mg/ml in water/buffer).
0814.11216 0150017
www.lifescience.roche.com
Table 3: Protease inhibitors offered by Roche
Diagnostics:
Inhibitors
Pack Size
Cat. No.
20 tablets
cpmplete Protease Inhibitor Cocktail Tablets (1 tablet for 50 ml)
3 × 20 tablets
11 697 498 001
11 836 145 001
cpmplete Mini (1 tablet for 10 ml)
25 tablets
11 836 153 001
cpmplete EDTA-free 
(1 tablet for 50 ml)
20 tablets
11 873 580 001
cpmplete Mini, EDTA-free
(1 tablet for 10 ml)
25 tablets
11 836 170 001
(4-Amidinophenyl)-methanesulfonyl fluoride (APMSF)
10 mg
10 917 575 001
Antipain-dihydrochloride
10 mg
11 004 646 001
10 mg
50 mg
100 mg
10 236 624 001
10 981 532 001
11 583 794 001
Bestatin
10 mg
50 mg
10 874 515 001
11 359 070 001
Calpain inhibitor I
25 mg
11 086 090 001
Calpain inhibitor II
25 mg
11 086 103 001
L-1-Chloro-3-(4-tosylamido)-7amino-2-heptanone-hydrochloride (TLCK)
100 mg
250 mg
10 874 485 001
10 874 493 001
L-1-Chloro-3-(4-tosylamido)-4phenyl-2-butanone (TPCK)
1g
10 874 507 001
Chymostatin
10 mg
11 004 638 001
3,4-Dichloroisocoumarin
10 mg
10 973 840 001
E-64
5 mg
10 mg
25 mg
11 585 673 001
10 874 523 001
11 585 681 001
2000 ATU
11 110 276 001
5 mg
25 mg
50 mg
100 mg
11 017 101 001
11 017 128 001
11 034 626 001
11 529 048 001
Aprotinin from bovine lung
Hirudin
Leupeptin
2-Macroglobulin from bovine
plasma
Pefabloc® SC (AEBSF)
Pefabloc® Plus
Pepstatin from Streptomyces species
Phenylmethylsulfonyl fluoride
(PMSF)
PMSF Plus
Phosphoramidon
Trypsin inhibitor from chicken egg
white
Trypsin inhibitor from soybean
25 inhibitor units
References
1
2
3
4
5
6
7
8
9
Umezawa, H. (1982) Annu. Rev. Microbiol. 36, 75.
Kassell, B. (1970) Methods Enzymol. 19, 844.
Umezawa, H. (1976) Methods Enzymol. 45, 678.
Hanada, K. et al. (1978) Agr. Biol. Chem. 42, 523.
Maniatis, T. et al. (1982) Molecular Cloning: A Laboratory Manual, 446, 
Cold Spring Harbor Laboratory, New York.
Markwardt, F. et al. (1973) Thromb. Res. 2, 343.
Schickaneder, E. et al. (1988) Fresenius Z. Anal. Chem. 330, 360.
North, M. J. (1989) in: Proteolytic enzymes — A practical approach p.117, 
Beynon, R. J. & Bond, J. S. (eds.), IRL-Press, Oxford.
Jazwinski, S. M. (1990) Methods Enzymol. 182, 154.
* available from Roche Diagnostics
Changes to 
previous version
Updated information regarding antipain-dihydrochloride specificity and suggested concentration.
Trademarks
COMPLETE is a trademark of ROCHE.
PEFABLOC is a trademarks of Pentypharm AG, Basel,
Switzerland.
All other product names and trademarks are the property
of their respective owners.
Regulatory 
Disclaimer
For life science research only. Not for use in diagnostic
procedures.
10 602 442 001
100 mg
500 mg
1g
11 429 868 001
11 585 916 001
11 429 876 001
100 mg
+ additive
1 g + additive
11 873 601 001
2 mg
10 mg
50 mg
10 253 286 001
11 359 053 001
11 524 488 001
1g
10 g
25 g
10 236 608 001
10 837 091 001
11 359 061 001
1 g + additive
11 873 636 001
11 873 628 001
5 mg
10 874 531 001
1g
10 109 878 001
50 mg
500 mg
10 109 886 001
10 109 894 001
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