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E-64, n-[n-( l-transepoxysuccinic acid)-l-leucyl]-agmatine, n-[n-(l-3-trans-carboxirane-2-carbonyl)-l-leucyl]-4-aminobutylguanidine, n-[n-(l-3-trans-carboxirane-2-carbonyl)-l-leucyl]-agmatine.
note: another name for l-3-trans-carboxyoxiran-2-carboxylic acid acid is l-transepoxysuccinic acid. agmatine is 4-aminobutylguanidine., trans-Epoxysuccinyl-L-leucylamido(4-guanidino)butane, L-trans-3-Carboxyoxiran-2-carbonyl-L-leucylagmatine, N-(trans-Epoxysuccinyl)-L-leucine 4-guanidinobutylamide
Empirical Formula (Hill Notation):
CAS Number:
Molecular Weight:
Numer MDL:
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Poziom jakości



>95% (HPLC)



masa cząsteczkowa

Mr 357.4


pkg of 10 mg (10874523001)
pkg of 25 mg (11585681001)



pH range



ethanol: water (1:1): soluble 20 mg/mL

temp. przechowywania


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The thiol protease inhibitor E-64 specifically inhibits papain and other cysteine proteases such as cathepsin B and L, bromelain, and ficin. Cathepsin A and D are not inhibited. L-lactate dehydrogenase from porcine heart is not inhibited, although the enzyme contains a functional thiol group.


E-64 is an effective ligand for affinity purification of cysteine proteases. When coupled to a thiolated affinity matrix, binding is no longer irreversible, but specificity is retained.
E-64 is used for the inhibition of papain and other cysteine proteases such as cathepsin B and L, bromelain, and ficin during isolation and purification of proteins and enzymes.
It has been used in the affinity chromatography for the purification of hemoglobin receptors.[1]
The inhibition of thiol proteases by E-64 appears to be of a non-competitive nature between the SH components. The inhibition is also irreversible, and is altered after gel filtration (Sephadex column) or dialysis after incubation of papain with E-64. The enzyme and inhibitor combine in an equimolar ratio.

Działania biochem./fizjol.

E-64 is an irreversible, potent, and highly selective cysteine protease inhibitor. E-64 does not react with the functional thiol group of non-protease enzymes, such as L-lactate dehydrogenase or creatine kinase. E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors, leupeptin and antipain. The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases, such as papain, actinidase, and cathepsins B, H, and L to form a thioether linkage. E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition, it is permeable in cells and tissues and has low toxicity.

Uwaga dotycząca przygotowania

Working concentration: 0.5 to 1.0 μg/ml (1.4 to 28.0 μM)
Working solution: Soluble to 20 mg/ml (stock solution) in a 1:1 (v/v) mixture of ethanol and water (vortexing or slight warming in water bath (40 °C) may facilitate dissolution). Also soluble in a neutral water/methanol solution, in water, methanol, acetic acid, pyridine and DMSO. Sparingly soluble in ethanol and propanol. Insoluble in acetone, chloroform, ethyl ether and benzene.
Storage conditions (working solution): -15 to -25 °C
Solutions are stable for one month when stored in aliquots at -15 to -25 °C.
E-64 is soluble in water. A 20 mg/ml solution can be prepared in water (heat may be needed). A suggested stock solution is a 1 mM aqueous solution. The effective concentration for use as a protease inhibitor is 1 to10 μM. Aqueous stock solutions are stable for months at -20 °C. Diluted solutions are stable for days at neutral pH. E-64 is stable from pH 2-10, but is unstable in ammonia or in HCl. E-64 is also soluble in DMSO, a 10 mM solution can be prepared in dry DMSO and stored at -20 °C. Subsequent dilutions were in culture medium. Solutions for injection were prepared by dissolving E-64 in 0.9% sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 0.9% sodium chloride (after adjusting the pH to 7.0 with acetic acid).


Soluble to 20 mg/ml in a 1:1 (v/v) mixture of ethanol and water. Solutions are stable for one month when stored in aliquots at -15 to -25 °C.

Inne uwagi

For life science research only. Not for use in diagnostic procedures.


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