產(chǎn)品詳情

  • 產(chǎn)品名稱：Caspase-10SubstrateAEVD-AFC

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  • 產(chǎn)品廠商：Biovision
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Caspase-10SubstrateAEVD-AFC
詳情介紹：
Sequence Ac-Ala-Glu-Val-Asp-AFC
Characteristics Ready-to-use fluorometric substrate for AEVD-dependent caspases. Activity of AEVD-dependent caspase activity can be quantified by fluorescent detection of free AFC after cleavage from the peptide substrate AEVD-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. Cell Lysis Buffer and other reagents used for caspase assays are also available separately.
Purity > 98?% by HPLC
Chemical Name Ac-AEVD-AFC, Caspase-10 Substrate, Fluorogenic
Formula C??H??F?N?O??
Permeability Not-permeable
Molecular Weight 685.6 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 AEVD-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.
8. Fold-increase in AEVD-dependent caspase 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
Product cited in: Baumgartner, Gerasimenko, Thorne, Ashurst, Barrow, Chvanov, Gillies, Criddle, Tepikin, Petersen, Sutton, Watson, Gerasimenko: "Caspase-8-mediated apoptosis induced by oxidative stress is independent of the intrinsic pathway and dependent on cathepsins." in: American journal of physiology. Gastrointestinal and liver physiology, Vol. 293, Issue 1, pp. G296-307, 2007 (PubMed).

Bai, Goodrich: "Different DNA lesions trigger distinct cell death responses in HCT116 colon carcinoma cells." in: Molecular cancer therapeutics, Vol. 3, Issue 5, pp. 613-9, 2004 (PubMed).