E-64
ROCHE/E64-RO - N-[N-(L-3-Trans-carboxirane-2-carbonyl)-L-leucyl]-agmatine
Synonym: E-64; n-[n-( l-transepoxysuccinic acid)
note: another name for l-
CAS Number: 66701-25-5
Empirical Formula (Hill Notation): C15H27N5O5
Molecular Weight: 357.41
MDL Number: MFCD00080261
Linear Formula: C15H27N5O5
Product Type: Chemical
assay | >95% (HPLC) |
form | powder |
InChI | 1S/C15H27N5O5/c1-8(2)7-9( |
InChI key | LTLYEAJONXGNFG-HBNTYKKESA |
manufacturer/tradename | Roche |
mol wt | Mr 357.4 |
packaging | pkg of 10 mg (10874523001) |
pkg of 25 mg (11585681001) | |
pH range | 2-10 |
Quality Level | 100 ![]() |
SMILES string | CC(C)C[C@H](NC(=O)[C@@H]1 |
solubility | ethanol: water (1:1): soluble 20 mg/mL |
storage temp. | 2-8°C |
Application: | 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. |
Application: | 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. 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. |
Biochem/physiol Actions: | 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 |
Other Notes: | For life science research only. Not for use in diagnostic procedures. |
Preparation Note: | 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. |
Preparation Note: | 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). |
Reconstitution: | 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. |
Specificity: | 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. |
Symbol | ![]() |
Signal word | Warning |
Hazard statements | H371 |
Precautionary statements | P260 - P264 - P270 - P308 + P311 - P405 - P501 |
RIDADR | NONH for all modes of transport |
WGK Germany | WGK 2 |
Flash Point(F) | does not flash |
Flash Point(C) | does not flash |
Purity | >95% (HPLC) |
Storage Temp. | 2-8°C |
UNSPSC | 12352200 |