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当前位置: 首页 > 产品中心 > Fluorescent_dyes > 生物素/Caspase-3 DEVD-R110荧光和比色分析试剂盒/30008-1/25分析
商品详细生物素/Caspase-3 DEVD-R110荧光和比色分析试剂盒/30008-1/25分析
生物素/Caspase-3 DEVD-R110荧光和比色分析试剂盒/30008-1/25分析
生物素/Caspase-3 DEVD-R110荧光和比色分析试剂盒/30008-1/25分析
商品编号: 30008-1
品牌: Biotium Inc
市场价: ¥2860.00
美元价: 1716.00
产地: 美国(厂家直采)
公司:
产品分类: 荧光染料
公司分类: Fluorescent_dyes
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
商品介绍
Caspase-3 is an active cell-death protease involved in the execution phase of apoptosis, during which cells undergo morphological changes such as DNA fragmentation, chromatin condensation and apoptotic body formation.

Caspase-3 is an active cell-death protease involved in the execution phase of apoptosis, during which cells undergo morphological changes such as DNA fragmentation, chromatin condensation and apoptotic body formation. Caspase-3 DEVD-R110 Fluorometric and Colorimetric Assay Kit provides a simple assay system for fast and highly sensitive detection of caspase-3 activity either by fluorescence or absorbance in mammalian cells. The fluorogenic and chromogenic substrate (Ac-DEVD)2-R110 contains two DEVD tetrapeptides and is completely hydrolyzed by the enzyme in two successive steps. Cleavage of the first DEVD peptide results in the monopeptide Ac-DEVD-R110 intermediate, which has absorption and emission wavelengths similar to those of R110 (λExEm = 496/520 nm) but has only about 10%of the fluorescence of the latter. Hydrolysis of the second DEVD peptide releases the dye R110, leading to a substantial fluorescence increase.

Although fluorometric detection of the end products is preferred because of the superior sensitivity, detection by absorbance is also possible. In fact, the extinction coefficient ofR110 is 10 times higher than that of p-nitroaniline (pNA), a dye commonly used in chromogenic substrates, making R110-based substrates significantly more sensitive than pNA-based substrates, even by colorimetric detection.

The assay kit includes Ac-DEVD-CHO, which is a caspase-3 inhibitor and can be used as a negative control. Also, R110 is provided in the kit for generating a standard curve, which can be used for quantifying caspase-3 activity.

Note: While caspase-3 preferentially cleaves the consensus sequence DEVD compared to other substrate sequences, other caspases also can cleave DEVD efficiently. Overlapping caspase substrate recognition limits the usefulness of caspase substrate peptides for distinguishing between different caspase activities in cell lysates.

Kit Components:

  • Cell lysis buffer
  • Assay buffer
  • Enzyme substrate (Ac-DEVD)2-R110
  • Enzyme inhibitor Ac-DEVD-CHO
  • R110

Features:

  • Fast: Fast enzyme kinetics.
  • Sensitive: The enzymatic reaction forms intensely yellow colored and highly green fluorescent rhodamine 110 (R110) product.
  • Versatile: Compatible with both fluorometric and colorimetric detection systems.

Biotium also offers the Caspase-3 DEVD-R110 Fluorometric HTS Assay Kit (catalog # 30009), a one step homogenous assay for high throughput screening (HTS) using fluorescent-based measurement. For real-time detection of caspase-3 activity in intact cells, see our novel NucView™488 Caspase-3 Substrate.

References

1. Cell Death Diff. 6, 99 (1999).

2. J Biol Chem 274, 11549 (1999).

3. J Biol Chem 275, 288 (2000).

4. Biochemistry, 38, 13906 (1999).

品牌介绍
★EB(溴化乙锭):溴化乙锭是一种高度灵敏的荧光染色剂,用于观察琼脂糖和聚丙烯酰胺凝胶中的DNA。溴化乙锭用标准302nm 紫外光透射仪激发并放射出橙红色信号,可用Polaroid 底片或带CCD 成像头的凝胶成像处理系统拍摄。 ★SYBR Green I/II是一种结合于所有dsDNA双螺旋小沟区域的具有绿色激发波长的染料。在游离状态下,SYBR Green I发出微弱的荧光,但一旦与双链DNA结合后,荧光大大增强。因此,SYBR Green I的荧光信号强度与双链DNA的数量相关,可以根据荧光信号检测出PCR体系存在的双链DNA数量。SYBR Green I 的最大吸收波长约为497nm,发射波长最大约为520nm。