You are viewing an incomplete version of our website. Please click to reload the website as full version.

Caspase-3 Substrate DEVD-AFC

Details for Product No. ABIN412342, Supplier: Log in to see
Enzyme Activity Assay (EAA), Functional Studies (Func)
Log in to see
Supplier Product No.
Log in to see

Showcase your results, aid the scientific community, and receive a full refund.

Contribute a validation

Learn more

Sequence Ac-Asp-Glu-Val-Asp-AFC
Purity > 98 % by HPLC
Molecular Weight 729 Da

Ready-to-use fluorometric substrate for caspase-3/CPP32 (Km = 9.7 µM) and related caspases that recognize the amino acid sequence DEVD. The sequence DEVD is based on caspase-3 cleavage site in poly (ADP-ribose) polymerase (PARP). CPP32 and related caspase activity can be quantified by fluorescent detection of free AFC after cleaved from the peptide substrate DEVD-AFC at Ex. = 400 nm and Em. = 505 nm, using a fluorometer or a 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.

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.
8. 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 Potect from light and moisture.
Storage -20 °C
Expiry Date 6-12 months
Product cited in: Pei, Yung, Yip et al.: "Desacyl ghrelin prevents doxorubicin-induced myocardial fibrosis and apoptosis via the GHSR-independent pathway." in: American journal of physiology. Endocrinology and metabolism, Vol. 306, Issue 3, pp. E311-23, 2014 (PubMed).

Chen, Ou, Li et al.: "Infection of pericytes in vitro by Japanese encephalitis virus disrupts the integrity of the endothelial barrier." in: Journal of virology, Vol. 88, Issue 2, pp. 1150-61, 2013 (PubMed).

Ren, Lu, Porollo et al.: "Tumor-suppressing function of caspase-2 requires catalytic site Cys-320 and site Ser-139 in mice." in: The Journal of biological chemistry, Vol. 287, Issue 18, pp. 14792-802, 2012 (PubMed).

Rui, Zhang, Xu et al.: "Reduction in IL-33 expression exaggerates ischaemia/reperfusion-induced myocardial injury in mice with diabetes mellitus." in: Cardiovascular research, Vol. 94, Issue 2, pp. 370-8, 2012 (PubMed).

Sémiramoth, Gleizes, Turbica et al.: "Afa/Dr-expressing, diffusely adhering Escherichia coli strain C1845 triggers F1845 fimbria-dependent phosphatidylserine externalization on neutrophil-like differentiated PLB-985 cells through an ..." in: Infection and immunity, Vol. 78, Issue 7, pp. 2974-83, 2010 (PubMed).

Griswold, Chang, Runko et al.: "Sir2 mediates apoptosis through JNK-dependent pathways in Drosophila." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 105, Issue 25, pp. 8673-8, 2008 (PubMed).

Lu, Caplan, Li et al.: "Polyunsaturated fatty acids block platelet-activating factor-induced phosphatidylinositol 3 kinase/Akt-mediated apoptosis in intestinal epithelial cells." in: American journal of physiology. Gastrointestinal and liver physiology, Vol. 294, Issue 5, pp. G1181-90, 2008 (PubMed).

Claud, Lu, Wang et al.: "Platelet-activating factor-induced chloride channel activation is associated with intracellular acidosis and apoptosis of intestinal epithelial cells." in: American journal of physiology. Gastrointestinal and liver physiology, Vol. 294, Issue 5, pp. G1191-200, 2008 (PubMed).

Mohammad, Goustin, Aboukameel et al.: "Preclinical studies of TW-37, a new nonpeptidic small-molecule inhibitor of Bcl-2, in diffuse large cell lymphoma xenograft model reveal drug action on both Bcl-2 and Mcl-1." in: Clinical cancer research : an official journal of the American Association for Cancer Research, Vol. 13, Issue 7, pp. 2226-35, 2007 (PubMed).

Feng, Bianchi, Sandmeyer et al.: "Bradykinin preconditioning improves the profile of cell survival proteins and limits apoptosis after cardioplegic arrest." in: Circulation, Vol. 112, Issue 9 Suppl, pp. I190-5, 2005 (PubMed).

Wesche-Soldato, Chung, Lomas-Neira et al.: "In vivo delivery of caspase-8 or Fas siRNA improves the survival of septic mice." in: Blood, Vol. 106, Issue 7, pp. 2295-301, 2005 (PubMed).

Yano, Itoh, Kubota et al.: "A prostacyclin analog prevents radiocontrast nephropathy via phosphorylation of cyclic AMP response element binding protein." in: The American journal of pathology, Vol. 166, Issue 5, pp. 1333-42, 2005 (PubMed).

Ray, Akbiyik, Bernstein et al.: "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).

Keedwell, Zhao, Hammond et al.: "A retinoid-related molecule that does not bind to classical retinoid receptors potently induces apoptosis in human prostate cancer cells through rapid caspase activation." in: Cancer research, Vol. 64, Issue 9, pp. 3302-12, 2004 (PubMed).

Jilling, Lu, Jackson et al.: "Intestinal epithelial apoptosis initiates gross bowel necrosis in an experimental rat model of neonatal necrotizing enterocolitis." in: Pediatric research, Vol. 55, Issue 4, pp. 622-9, 2004 (PubMed).

Suh, Mutoh, Nagashima et al.: "The organellular chloride channel protein CLIC4/mtCLIC translocates to the nucleus in response to cellular stress and accelerates apoptosis." in: The Journal of biological chemistry, Vol. 279, Issue 6, pp. 4632-41, 2004 (PubMed).

Sherwood, Sternberg: "Anchor cell invasion into the vulval epithelium in C. elegans." in: Developmental cell, Vol. 5, Issue 1, pp. 21-31, 2003 (PubMed).

Nowak: "Protein kinase C-alpha and ERK1/2 mediate mitochondrial dysfunction, decreases in active Na+ transport, and cisplatin-induced apoptosis in renal cells." in: The Journal of biological chemistry, Vol. 277, Issue 45, pp. 43377-88, 2002 (PubMed).

Cummings, Schnellmann: "Cisplatin-induced renal cell apoptosis: caspase 3-dependent and -independent pathways." in: The Journal of pharmacology and experimental therapeutics, Vol. 302, Issue 1, pp. 8-17, 2002 (PubMed).

Hall, Matter, Wang et al.: "Hyperglycemia inhibits vascular smooth muscle cell apoptosis through a protein kinase C-dependent pathway." in: Circulation research, Vol. 87, Issue 7, pp. 574-80, 2000 (PubMed).