Caspase-1 Substrate YVAD-AFC

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Enzyme Activity Assay (EAA), Functional Studies (Func)
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Sequence Ac-Tyr-Val-Ala-Asp-AFC
Characteristics Ready-to-use fluorometric substrate for caspase-1/ICE and related caspases that recognize the amino acid sequence YVAD. Caspase-1 and related caspase activity can be quantified by fluorescent detection of free AFC after cleaved from the peptide substrate YVAD-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. The ready-to-use caspase substrate provides an economic alternative for researchers who perform large volume caspase assays. Cell Lysis Buffer, and DTT used for caspase assays are also available separately.
Purity > 99 % by HPLC
Chemical Name Ac-YVAD-AFC, Caspase-1 Substrate, Fluorogenic
Formula C₃₃H₃₆F₃N₅O₁₀
Permeability Not-permeable
Molecular Weight 719.7 g/mol
Protocol 1. Induce apoptosis or treat cells by desired method. Concurrently incubate a control culture without treatment. Note: Active recombinant human caspase-1 is available to use as a positive control and incubate at 37 °C for 1-2 hour.
7. Read samples in a fluorometer equipped with a 400-nm excitation and 505-nm emission filters. For a plate-reading set-up, transfer the samples to a 96-well plate. You may also perform the entire assay directly in a 96-well plate. Fold-increase in YVAD-dependent caspase activity can be determined by comparing these results with the level of the untreated 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: Ray, Akbiyik, Bernstein, Phipps: "CD40 engagement prevents peroxisome proliferator-activated receptor gamma agonist-induced apoptosis of B lymphocytes and B lymphoma cells by an NF-kappaB-dependent mechanism." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 174, Issue 7, pp. 4060-9, 2005 (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).

Coletti, Yang, Marazzi, Sassoon: "TNFalpha inhibits skeletal myogenesis through a PW1-dependent pathway by recruitment of caspase pathways." in: The EMBO journal, Vol. 21, Issue 4, pp. 631-42, 2002 (PubMed).