胱天蛋白酶Caspase抑制剂 FAM-VAD-FMK
Ex (nm) | 493 | Em (nm) | 517 |
分子量 | 691.66 | 溶剂 | DMSO |
存储条件 | 在零下15度以下保存, 避免光照 |
胱天蛋白酶Caspase抑制剂 FAM-VAD-FMK是美国AAT Bioquest生产的用于Caspase的试剂,FAM-VAD是一种绿色荧光细胞渗透性polycaspase抑制剂靶向胱天蛋白酶1,2,3,6,8,9或10。一旦进入细胞,所述抑制剂共价结合于胱天蛋白酶活性,其产生绿色荧光。当加入到细胞群体中,FAM-VAD-FMK探针进入每个细胞和共价结合到驻留在胱天蛋白酶活性的异二聚体的大亚基,从而抑制进一步的酶活性的反应性半胱氨酸残基。结合的标记试剂被保留在细胞内,而任何未结合的试剂将扩散出细胞,并冲走。绿色荧光信号的直接测量活性胱天蛋白酶的存在于细胞中的试剂加入的时间量。含有结合的标记试剂细胞可以通过96 - 孔板的基于荧光,荧光显微镜或流式细胞仪进行分析。百萤生物是AAT Bioquest的中国代理商,为您提供优质的胱天蛋白酶Caspase抑制剂 FAM-VAD-FMK。
样品实验方案
溶液制备
1.储备溶液
所有未使用的储备溶液应分为一次性使用的等分试样,并在制备后储存在-20°C下。 避免重复冻融循环。
1.1 FAM-VAD-FMK储备溶液:
在DMSO中准备2-5 mM的储备溶液。
样品操作及分析
1.根据需要处理细胞。
2.通过用20 mM Hepes缓冲液(HHBS)在Hanks中稀释DMSO储备溶液,制备2X渗透性半胱天冬酶底物(20 µM)分析溶液。
3.将等体积的处理过的细胞与2X半胱天冬酶底物测定溶液混合,并在37°C,5%CO2的培养箱中孵育细胞至少1小时。
4.用HHBS清洗细胞至少一次。
5.通过流式细胞仪,荧光显微镜或荧光酶标仪监测荧光强度。
参考文献
Suppression of human T cell proliferation by the caspase inhibitors, z-VAD-FMK and z-IETD-FMK is independent of their caspase inhibition properties
Authors: Lawrence CP, Chow SC.
Journal: Toxicol Appl Pharmacol. (2012)
[Inhibition of elicitation of allergic contact dermatitis by topical use of Z-VAD-FMK, a broad caspase inhibitor: experiment in mice]
Authors: Li YY, Yan CL.
Journal: Zhonghua Yi Xue Za Zhi (2012): 1992
Intracochlear perfusion of leupeptin and z-VAD-FMK: influence of antiapoptotic agents on gunshot-induced hearing loss
Authors: Abaamrane L, Raffin F, Schmerber S, Sendowski I.
Journal: Eur Arch Otorhinolaryngol (2011): 987
Plasmodium falciparum metacaspase PfMCA-1 triggers a z-VAD-fmk inhibitable protease to promote cell death
Authors: Meslin B, Beavogui AH, Fasel N, Picot S.
Journal: PLoS One (2011): e23867
Structure of human caspase-6 in complex with Z-VAD-FMK: New peptide binding mode observed for the non-canonical caspase conformation
Authors: Muller I, Lamers MB, Ritchie AJ, Dominguez C, Munoz-Sanjuan I, Kiselyov A.
Journal: Bioorg Med Chem Lett (2011): 5244
Pretreatment with pancaspase inhibitor (Z-VAD-FMK) delays but does not prevent intraperitoneal heat-killed group B Streptococcus-induced preterm delivery in a pregnant mouse model
Authors: Equils O, Moffatt-Blue C, Ishikawa TO, Simmons CF, Ilievski V, Hirsch E.
Journal: Infect Dis Obstet Gynecol (2009): 749432
[Attenuation of allergic contact dermatitis by Z-VAD-FMK, a broad caspase inhibitor: experiment with mice]
Authors: Li YY, Yan CL, Xu W.
Journal: Zhonghua Yi Xue Za Zhi (2008): 3153
Bodipy-VAD-Fmk, a useful tool to study yeast peptide N-glycanase activity
Authors: Witte MD, Descals CV, de Lavoir SV, Florea BI, van der Marel GA, Overkleeft HS.
Journal: Org Biomol Chem (2007): 3690
Caspase inhibitor Z-VAD-FMK enhances the freeze-thaw survival rate of human embryonic stem cells
Authors: Heng BC, Clement MV, Cao T.
Journal: Biosci Rep (2007): 257
Effects of caspase inhibitors (z-VAD-fmk, z-VDVAD-fmk) on Nile Red fluorescence pattern in 7-ketocholesterol-treated cells: investigation by flow cytometry and spectral imaging microscopy
Authors: Vejux A, Lizard G, Tourneur Y, Riedinger JM, Frouin F, Kahn E.
Journal: Cytometry A (2007): 550
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