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  • 产品名称：Caspase-4SubstrateLEVD-AFC

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  • 产品厂商：Biovision
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简单介绍：
Caspase-4SubstrateLEVD-AFC
详情介绍：
Sequence Ac-Leu-Glu-Val-Asp-AFC
Characteristics Ready-to-use fluorometric substrate for caspase-4 and related caspases that recognize the amino acid sequence LEVD. Caspase-4 and related caspase activity can be quantified by fluorescent detection of free AFC after cleavage from the peptide substrate LEVD-AFC at Ex. = 400 nm and Em. = 505 nm, using a fluorometer or multi-well fluorescence plate reader. Alternatively, a shift in fluorescence from blue to green upon cleavage can be visualized using a hand-held long-UV lamp.
Purity > 99 % by HPLC
Chemical Name Ac-LEVD-AFC, Caspase-4 Substrate, Fluorogenic
Formula C₃₂H₄₀F₃N₅O₁₁
Permeability Not-permeable
Molecular Weight 727.5 g/mol
Protocol 1. Induce apoptosis in cells by desired method. Concurrently incubate a control culture without induction.
2. Count cells and pellet 1-5 x 106 cells or use 50-200 µg cell lysates if protein concentration has been measured.
3. Resuspend cells in 50 µL of chilled Cell Lysis Buffer containing 10 mM DTT to each sample.
6. Add 5 µL of the 1 mM DEVD-AFC (50 µ M final conc.) into each tube individually and incubate at 37 °C for 1-2 hour.
7. Read samples in a fluorometer equipped with a 400-nm excitation filter and 505-nm emission filter. For a plate-reading set-up, transfer the samples to a 96-well plate. You may perform the entire assay directly in a 96-well plate. Fold-increase in caspase-3 activity can be determined by comparing these results with the level of the uninduced control.
Restrictions For Research Use only
Format Liquid
Handling Advice Protect from light and moisture
Storage -20 °C
Expiry Date 12 months