E 64d
![mRNA synthesis](/media/diy/images/page/figure1-mrna.png)
mRNA synthesis
In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
![Tyramine Signal Amplification (TSA)](/media/diy/images/page/figure2-01.png)
Tyramide Signal Amplification (TSA)
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
![screening library](/media/diy/images/page/figure3-01.png)
Phos Binding Reagent Acrylamide
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
![Cell Counting Kit-8 (CCK-8)](/media/diy/images/page/CCK-8.jpg)
Cell Counting Kit-8 (CCK-8)
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
![SYBR Safe DNA Gel Stain](/media/diy/images/page/SYBR Safe DNA Gel Stain.png)
SYBR Safe DNA Gel Stain
Safe and sensitive stain for visualization of DNA or RNA in agarose or acrylamide gels.
![Inhibitor Cocktails](/media/diy/images/page/Inhibitor Cocktails.jpg)
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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- 4. Dziekan, Jerzy Michal. "Molecular analysis of antimalarial therapies : from drug development to mode of action." Nanyang Technological University. 2019.
- 5. Maxine Bauzon, Penelope M. Drake, et al. "Maytansine-bearing antibody-drug conjugates induce in vitro hallmarks of immunogenic cell death selectively in antigen-positive target cells." OncoImmunology.Received 09 Aug 2018, Accepted 12 Dec 2018, Published online: 22 Jan 2019.
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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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