Caspase-6 Fluorogenic Substrate
Quick Overview for Caspase-6 Fluorogenic Substrate (ABIN924942)
应用范围
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原理
- Peptide substrate labeled at the carboxy end with AFC (7-amino-4-trifluoromethyl coumarin). Designed to measure Caspase-6 activity in vitro.
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序列
- Ac-Val-Glu-Ile-Asp-AFC, Ac-VEID-AFC
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特异性
- Highly specific substrate for Caspase-6. May be weak substrate for Caspases-1 and -3.
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应用备注
- For in vitro assays of Caspase-6 activity. Can be used with purified or partially purified enzymes or possibly with crude cell lysates (if the Caspase-6 Inhibitor is included to determine background protease activity).
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说明
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Fluorometer calibration: The fluorometer is calibrated using known concentrations of free AFC (Excitation = 400 nm, Emission = 505 nm) to generate a standard curve of fluorescence versus µ-moles AFC. Samples: Can be either purified or partially purified enzyme preparations. Application to crude cell lysates has not been confirmed. If crude cell lysates are to be assayed, the non- specific protease background must be determined using our Caspase-6 Inhibitor.
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限制
- 仅限研究用
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储存条件
- -20 °C
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背景
- Peptide substrate labeled at the carboxy end with AFC (7-amino-4-trifluoromethyl coumarin). Designed to measure Caspase-6 activity in vitro. Caspase-6 (also known as Mch2) is a member of the caspase family of cysteine proteases involved in apoptosis. It is a member of the Group III caspases (6, 8, and 9) which prefer the (L/V)EXD sequence as a substrate. Caspase-6 prefers a hydrophobic amino acid at P4, along with caspases-1 and -4, as opposed to the preference for Asp seen with caspases-2, -3, and -7. This is at odds with the gene sequence alignment that predicts Caspase-6 is more closely related to caspases-3 and -7 than to caspase-1. The preference by Caspase-6 for beta-branched amino acids in P4 fits well with the one known natural substrate, lamin A, and distinguishes it from caspases-1 and -4. Reconstitution experiments indicate that Caspase-6 activates caspases-3 and -7 and is therefore part of the proteolytic cascade that initiates apoptosis.
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分子量
- 727.69 Da
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