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  • 產品名稱：Caspase-4SubstrateLEVD-pNA

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  • 產品廠商：Biovision
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Caspase-4SubstrateLEVD-pNA
詳情介紹：
Sequence Ac- Leu-Glu-Val-Asp-pNA
Characteristics Ready-to-use colorimetric substrate for caspase-4 and related caspases that recognize the amino acid sequence LEVD. Caspase-4 and related caspase activity can be quantified by spectrophotometric detection of free pNA (l= 400 nm) after cleavage from the peptide substrate LEVD-pNA, using a spectrophotometer or multi-well plate reader. The ready-to-use caspase substrate provides an economic alternative for large volume users.
Purity > 98?% by HPLC
Chemical Name Ac-LEVD-pNA, Caspase-4 Substrate, Colorimetric
Formula C??H??N?O??
Permeability Not-permeable
Molecular Weight 636.65 g/mol
Protocol 1. Induce apoptosis in cells by desired method. Concurrently incubate a control culture without induction. Note: Active recombinant human caspase-4 is available to use as a positive control to a fresh tube and put on ice.
6. Assay protein concentration.
7. Dilute 100-300 μg protein to 50 μL Cell Lysis Buffer for each assay.
8. Add 50 μL of 2X Reaction Buffer containing 10 mM DTT to each sample.
9. Add 5 μL of the 4 mM of LEVD- p NA (200 μM final conc.) and incubate at 37 °C for 1-2 hour.
10. Read samples at 400- or 405-nm in a microtiter plate reader, or spectrophotometer using a 100- μL micro quartz cuvet (Sigma), or dilute sample to 1 mL with Dilution Buffer and using regular cuvet (note: Dilution of the samples proportionally decreases the reading). You may also perform the entire assay directly in a 96-well plate. Fold-increase in caspase activity can be determined by comparing these results with the level of the uninduced control. Note: Background reading from cell lysates and buffers should be subtracted from the readings of both induced and the uninduced samples before calculating fold increase in caspase activity.
Restrictions For Research Use only
Format Liquid
Handling Advice Protect from light and moisture
Storage -20 °C
Expiry Date 12 months