E 64d
mRNA synthesis
In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
Tyramide Signal Amplification (TSA)
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Phos Binding Reagent Acrylamide
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
Cell Counting Kit-8 (CCK-8)
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
SYBR Safe DNA Gel Stain
Safe and sensitive stain for visualization of DNA or RNA in agarose or acrylamide gels.
Inhibitor Cocktails
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
E-64d是E-64c的膜通透性衍生物和巯基蛋白酶的抑制剂，被用于检测其对完整血小板中钙蛋白酶（calpain）的抑制活性。
E-64c或E-64d均可以抑制钙蛋白酶的活性。将完整血小板与抑制剂孵育10分钟，在裂解前洗去细胞外的抑制剂，E-64c和leupeptin均不能抑制蛋白水解，但E-64d可以。E-64d能够进入血小板，不会通过洗涤而除去。然而，E-64c和E-64d均能够抑制裂解血小板中的蛋白酶活性。由于该蛋白酶的活性是Ca2+依赖型的，而且能够被钙蛋白酶抑制剂抑制，因而被认为是钙蛋白酶。E-64d能够进入完整的血小板：i)洗涤除去细胞外的抑制剂，裂解血小板后检测不到蛋白酶活性；ii)与E-64d而非E-64c孵育，可以激活血小板，从而抑制由A23187和钙激活的完整血小板中钙蛋白酶的水解活性。
参考文献：
1. M. Tamai, K. Matsumoto, S. Omura, I. Koyama, Y. Ozawa, K. Hanada J. Pharmacobio-Dyn., 9 (1986), pp. 672–677.
2. E. B. McGowan, E. Becker, and T. C. Detwiler, INHIBITION OF CALPAIN IN INTACT PLATELETS BY THE THIOL PROTEASE INHIBITOR E-64d. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS , Vol. 158, No. 2, 1989.
3. Carmen JC, Sinai AP. The Differential Effect of Toxoplasma Gondii Infection on the Stability of BCL2-Family Members Involves Multiple Activities. Front Microbiol. 2011 Jan 24; 2:1.
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| Physical Appearance | A solid |
| Storage | Store at -20°C |
| M.Wt | 342.43 |
| Cas No. | 88321-09-9 |
| Formula | C17H30N2O5 |
| Solubility | insoluble in H2O; ≥17.12 mg/mL in DMSO; ≥18.5 mg/mL in EtOH |
| Chemical Name | ethyl (2S,3S)-3-[[(2S)-4-methyl-1-(3-methylbutylamino)-1-oxopentan-2-yl]carbamoyl]oxirane-2-carboxylate |
| SDF | Download SDF |
| Canonical SMILES | CCOC(=O)C1C(O1)C(=O)NC(CC(C)C)C(=O)NCCC(C)C |
| 运输条件 | 蓝冰运输或根据您的需求运输。 |
| 一般建议 | 不同厂家不同批次产品溶解度各有差异，仅做参考。若实验所需浓度过大至产品溶解极限，请添加助溶剂助溶或自行调整浓度。溶液形式一般不宜长期储存，请尽快用完。 |
| 细胞实验[1]： | |
|
细胞系 |
血小板 |
|
溶解方法 |
在DMSO中的溶解度 >10 mM。为了获得更高的浓度，可以将离心管在37°C加热10分钟和/或在超声波浴中震荡一段时间。原液可以在-20°C以下储存几个月。 |
|
反应条件 |
50 μg/mL；10 min |
|
应用 |
A23187和钙可以共同激活血小板，它们也可以导致calpain催化的ABP和talin蛋白水解。在缺乏抑制剂时，A23187导致ABP和talin的完全降解。E 64d是细胞通透性的抑制剂，抑制细胞内的蛋白水解，有些抑制在最低测试浓度 20 μg/mL 时观察到，而50 μg/mL 浓度的E 64d基本可以完全抑制蛋白水解。 |
| 动物实验[2]： | |
|
动物模型 |
雄性Sprague-Dawley大鼠 |
|
剂量 |
4 μg；腹腔注射 |
|
应用 |
E 64d治疗后，小鼠用penicillin处理诱导癫痫反复发作。结果表明，E 64d显著减少在齿状回颗粒上层和海马CA3区域的异常苔藓纤维发芽。在无E 64d治疗大鼠中，在齿状回和CA3区域的锥体层有明显的苔藓纤维末端聚集。而在E 64d治疗大鼠中，在齿状回颗粒上层和CA3区域中的苔藓纤维末端聚集显著减少。 |
|
注意事项 |
请测试所有化合物在室内的溶解度，实际溶解度和理论值可能略有不同。这是由实验系统的误差引起的，属于正常现象。 |
|
References: [1] McGowan E B, Becker E, Detwiler T C. Inhibition of calpain in intact platelets by the thiol protease inhibitor E-64d. Biochemical and biophysical research communications, 1989, 158(2): 432-435. [2] Ni H, Ren S, Zhang L, et al. Expression profiles of hippocampal regenerative sprouting-related genes and their regulation by E-64d in a developmental rat model of penicillin-induced recurrent epilepticus. Toxicology letters, 2013, 217(2): 162-169. |
|
| E-64d (合成的E-64的类似物、E-64c的乙基酯)是一个不可逆的、膜通透性的溶酶体和胞浆半胱氨酸蛋白酶抑制剂，抑制钙蛋白酶（calpain）和半胱氨酸蛋白酶cathepsins F、K、B、H和L。. | ||||||
| 靶点 | cathepsins F | cathepsins K | cathepsins B | cathepsins H | cathepsins L | |
| IC50 | ||||||
质量控制和MSDS
- 批次：
化学结构
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