E-64-Proteinase-inhibitor-E-64-DataSheet-生命科学试剂-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEE-64Cat. No.: HY-15282CAS No.: 66701-25-5Synonyms: Proteinase inhibitor E 64分式: CHNO分量: 357.41作靶点: Cathepsin; Autophagy作通路: Metabolic Enzyme/Protease; Autophagy储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 mon

2、th溶解性数据体外实验 DMSO : 50 mg/mL (139.90 mM; Need ultrasonic)H2O : 36 mg/mL (100.72 mM; Need ultrasonic and warming)Mass Solvent1 mg 5 mg 10 mg Concentration制备储备液1 mM 2.7979 mL 13.9895 mL 27.9791 mL5 mM 0.5596 mL 2.7979 mL 5.5958 mL10 mM 0.2798 mL 1.3990 mL 2.7979 mL请根据产品在不同溶剂中的溶解度，选择合适的溶剂配制储备液，并请注意储备液的保

3、存式和期限。体内实验请根据您的实验动物和给药式选择适当的溶解案，配制前请先配制澄清的储备液，再依次添加助溶剂(为保证实验结果的可靠性，体内实验的作液，建议您现现配，当天使；澄清的储备液可以根据储存条件，适当保存；以下溶剂前的百分 指该溶剂在您配制终溶液中的体积占)：1. 请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.75 mg/mL (7.69 mM); Clear solution2. 请依序添加每种溶剂： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.

4、75 mg/mL (7.69 mM); Clear solution1/3 Master of Small Molecules 您边的抑制剂师www.MedChemE3. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.75 mg/mL (7.69 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 E-64种有效的不可逆的半胱氨酸蛋酶 (cysteine proteases) 抑制剂，抑制 papain 的 IC50 为 9 nM。IC50 & Target IC50: 9 nM (Papain) 1体外研究 E-6

5、4 is a cathepsin B-specific inhibitor, and its binding modes with papain, actinidin, cathepsin L, andcathepsin K have been reviewed at the atomic level. E-64 has been widely used as a potent and irreversible(covalent-type) inhibitor for many cysteine proteases such as papain, ficin, actinidin, cathe

6、psin B and L 1.The S.cervi adult parasites are incubated in the Krebs Ringer bicarbonate (KRB) maintenance medium for 8h at 37C, 5% CO2 with 5, 10, 20 and 40 M concentration of E-64. E-64 shows a concentration and timedependent decrease in motility and viability of the parasites (EC50=16 M) 2.体内研究 A

7、 broad spectrum of expression of CD4 and CD19 is found exists in both the islets and pancreatic lymphnodes (PLNs) and that anti-serpin B13 mAb exposure causes a significant shift that favored cells expressinglow-to-intermediate amounts of these markers. However, this shift is abolished in animals th

8、at receive anti-serpin B13 mAb in the presence of the protease inhibitor E-64 (E64), which maintains its blocking activityunder the experimental conditions used 3. Dahl salt-sensitive (SS) rats are fed an 8% high salt NaCl dietand intravenously infused with the irreversible cysteine cathepsin inhibi

9、tor E-64 (1 mg/day) or the vehicle(control). Both the control and E-64 infused groups develope significant hypertension and kidney damage,and no difference of the mean arterial pressure and the hypertension-associated albuminuria is observedbetween the groups 4.PROTOCOLKinase Assay 2 The Cathepsin B

10、 activity is determined using Z-Arg-Arg-4mNA as substrate with slight modifications. Thecrude extract is pre-incubated at 37C for 5 min in 50 mM sodium acetate buffer, pH 5.0 containing 1 mMEDTA and 5 mM DTT. The substrate (final concentration, 100 M) is added to make the final assay volumeof 1 mL.

11、The reaction mixture is incubated at 37C for 30 min. The reaction is terminated by adding equalvolume of stopping reagent containing Fast Garnet GBC salt (1 mg/mL), 10 mM pHMB and 50 mM EDTA, pH6.0. The extraction of product, -napthylamine (-NA), is carried out with n-butanol. After complete layerse

12、paration, the absorbance is measured in n-butanol layer and activity is calculated using molar extinctioncoefficient of -napthylamine solution as 31.5 M/cm per sec at 520 nm. One unit of enzyme activity is definedas the amount of enzyme liberating 1 mol of NA per minute at 37C. The half maximal inhi

13、bitoryconcentration (IC50) is calculated by plotting the graph between the different concentration of E-64 and the% inhibition in cathepsin B activity. Here, IC50 indicates the concentration of the E-64 required to inhibit theparasitic cathepsin B activity by half 2.MCE has not independently confirm

14、ed the accuracy of these methods. They are for reference only.2/3 Master of Small Molecules 您边的抑制剂师www.MedChemEAnimal Mice 3Administration 34 The NOD/LtJ and BDC2.5 T cell receptor (TCR) transgenic NOD mice are used to study the effects oftreatment with anti-serpin B13 monoclonal antibody (mAb). Fou

15、r-week-old female NOD/LtJ mice are injectedintravenously four times over a period of 10 days with anti-serpin B13 mAb (100 g/injection). In addition,during the same period, some animals are also injected intraperitoneally with the protease inhibitor E64 at 10mg/kg/day for several days. Control mice

16、are treated with diluent (a sterilized PBS solution containing 10%DMSO) and control IgG. The solutions containing E64 or DMSO are prepared immediately before use.Twenty-four hours after the last injection, the mice are killed, and cells from their lymphoid organs andpancreatic islets are subjected t

17、o FACS analysis.Rats 4Seven-week old male Dahl Salt Sensitive rats (SS/JrHsdMcwi) are used. Briefly, 8-week old anesthetized SSrats have their left femoral artery and vein catheterized. Both catheters are fixed and exteriorized from theback of the neck and the arterial line is connected to a heparin

18、ized saline infusion pump that is in line with ablood pressure transducer, and the venous line is connected to a saline infusion pump. Animals are allowed360 movement using a tether-swivel system. This preparation allowed chronic venous infusion and arterialblood pressure measurement in conscious, f

19、reely moving rats. A stable baseline blood pressure is obtainedfor 4 days prior to switching both groups to an 8.0% NaCl diet and the simultaneous addition of E-64 (1mg/day; 280 mM stock in DMSO) or the vehicle (DMSO in saline) control to the venous catheter. Daily MAPis calculated by averaging MAP

20、taken every min over the beginning 3 h period of the rat sleep cycle.MCE has not independently confirmed the accuracy of these methods. They are for reference only.户使本产品发表的科研献 LWT-FOOD SCI TECHNOL. 2018 Jan, 87; 186-193 Int J Oncol. 2019 May. Dev Comp Immunol. 2018 Jan;78:114-123. J Aquat Food Prod T. 2019 Jul.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Matsumoto K, et al. Structural basis of inhibition of cysteine proteases by E