| Product Name | Renin FRET Substrate IDABCYL - g - Abu - IHPFHLVIHT - EDANS |
| Size | 1 mg |
| Catalog # | AS-24478 |
| US$ | $227 |
DABCYL-GABA-Ile-His-Pro-Phe-His-Leu-Val-Ile-His-Thr-EDANS (also called Renin Substrate I in some literature) contains a renin cleavage site that occurs in the N-terminal peptide of human angiotensinogen. Cleavage of the substrate occurs specifically at the Leu-Val bond and corresponds to the renin cleavage site of angiotensinogen. This fluorogenic peptide substrate is used to continuously measure the proteolytic activity of human renin. The assay relies upon FRET-mediated, intramolecular fluorescence quenching that occurs in the intact peptide substrate. Efficient fluorescence quenching occurs as a result of favorable energetic overlap of the EDANS excited state and the DABCYL absorption, and the relatively long excited state lifetime of the EDANS fluorophore. Cleavage of the substrate by renin liberates the peptidyl-EDANS fragment from proximity with the DABCYL acceptor, restoring the fluorescence of the EDANS moiety. This leads to a time-dependent increase in fluorescence intensity, directly related to the extent of substrate cleavage. The kinetics of renin-catalyzed hydrolysis of this substrate is shown to be consistent with a simple substrate inhibition model with a substrate Km approximately equal to 1.5 mM at physiological pH. The substrate can detect renin as low as 30 ng/mL after an incubation of only 3-5 min. Minimum Purity: 95% | |
| Detailed Information |  Material Safety Data Sheets (MSDS) |
| Storage | -20°C desiccated and protected from light |
| References | Wang GT, et al. (1993). A continuous fluorescence assay of renin activity. Anal Biochem 210, 351-9, Nakamura N, et al. (1991). Identification of the active site of human renin with use of new fluorogenic peptides. J Biochem (Tokyo) 109, 741-5, Murakami K, et al. (1981). New fluorogenic substrates for renin. Anal Biochem 110, 232-9. |
| Molecular Weight | 1797.9 |
| DABCYL-g-Abu-IHPFHLVIHT-EDANS | |
| Sequence(Three-Letter Code) | DABCYL - g - Abu - Ile - His - Pro - Phe - His - Leu - Val - Ile - His - Thr - EDANS |
| Spectral Properties | Abs/Em = 355/500 nm |
| Solvent System | DMSO |
| Product Citations | Wu, Zhongren. et al. BMC Biochemistry. 9 (2008). |
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《血管紧张素转换酶抑制剂在心血管病中应用的中国专家共识》.PDF(242.69k)
1.对于抑制剂筛选工作（求ic50）是不是体系内酶与底物的量（底物应该是过量的）对实验结果影响不大。
2.如果要求算Km值，是不是需要知道反应产物的绝对量。反应时间文献上都是5分钟，反应速度就用反应产物量除以反应时间即可。
3.酶是进口分装的，规格5U，一次用不完，用PBS稀释后如何保存
谢谢
2、可逆抑制剂：包括
a、竞争性抑制剂，抑制剂与底物竞争性结合酶反应中心，使Km增大，而Vmax不变 b、非竞争性抑制剂，酶与抑制剂结合后还能与底物结合，但活性降低，使Vmax减小，而Km不变
c、反竞争性抑制剂，酶只能与底物结合后才能与抑制剂结合，Vmax与Km都减小
可逆抑制剂可用透析等方法除去，使酶恢复作用
1、测定酶比活力：底物需要过量么？测定时间多长？是否可以加入过量的底物，然后测定3min吸光度的增加值，从吸光度的变化值计算比活力。
2、在酶抑制剂筛选的过程中，是否需要保证底物过量？还是要水浴一定时间让反应完全？我看到文献说用终浓度为0.1μmol/mL的底物，终浓度为45U/ml的酶，我想问，假设总体积为1ml，那么终浓度为45U/ml的酶岂不是每分钟能转化4.8μmol的底物？那么0.1μmol/mL的底物不就几秒钟就反应完了？那么怎么测定初速度？
