產(chǎn)品詳情
簡單介紹:
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). |