Caspase-9 Substrate LEHD-AFC

Details for Product No. ABIN412395
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Application
Functional Studies (Func), Enzyme Activity Assay (EAA)
Pubmed 19 references available
Catalog no. ABIN412395
Quantity 200 tests
Price
280.50 $   Plus shipping costs $45.00
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Sequence Ac-Leu-Glu-His-Asp-AFC
Characteristics Ready-to-use fluorometric substrate for caspase-9/Mch-6 and related caspases that recognize the amino acid sequence LEHD. Caspase activity can be quantified by fluorescent detection of free AFC after cleavage from the peptide substrate LEHD-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 amount of caspase assays. Cell Lysis Buffer for caspase assays are also available separately.
Purity > 98 % by HPLC
Molecular Weight 765.7 Da
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 LEHD-AFC (50 µM 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. Fold-increase in LEHD-dependent 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: Ding, Lin, McGill et al.: "Essential role for caspase-8 in transcription-independent apoptosis triggered by p53." in: The Journal of biological chemistry, Vol. 275, Issue 49, pp. 38905-11, 2001 (PubMed).

Tabata, Tabata, Grabowski et al.: "Roles of NF-kappaB and 26 S proteasome in apoptotic cell death induced by topoisomerase I and II poisons in human nonsmall cell lung carcinoma." in: The Journal of biological chemistry, Vol. 276, Issue 11, pp. 8029-36, 2001 (PubMed).

Vu, Bortner, Cidlowski: "Differential involvement of initiator caspases in apoptotic volume decrease and potassium efflux during Fas- and UV-induced cell death." in: The Journal of biological chemistry, Vol. 276, Issue 40, pp. 37602-11, 2001 (PubMed).

Coletti, Yang, Marazzi et al.: "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).

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).

Ghatak, Misra, Toole: "Hyaluronan oligosaccharides inhibit anchorage-independent growth of tumor cells by suppressing the phosphoinositide 3-kinase/Akt cell survival pathway." in: The Journal of biological chemistry, Vol. 277, Issue 41, pp. 38013-20, 2002 (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).

Kawakami, Kawakami, Puri: "Intratumor administration of interleukin 13 receptor-targeted cytotoxin induces apoptotic cell death in human malignant glioma tumor xenografts." in: Molecular cancer therapeutics, Vol. 1, Issue 12, pp. 999-1007, 2002 (PubMed).

Cummings, McHowat, Schnellmann: "Role of an endoplasmic reticulum Ca2+-independent phospholipase A2 in cisplatin-induced renal cell apoptosis." in: The Journal of pharmacology and experimental therapeutics, Vol. 308, Issue 3, pp. 921-8, 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).

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).

Lieman, Worley, Harbour: "Loss of Rb-E2F repression results in caspase-8-mediated apoptosis through inactivation of focal adhesion kinase." in: The Journal of biological chemistry, Vol. 280, Issue 11, pp. 10484-90, 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).

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).

Wei, Wang, Topczewski et al.: "Saturated fatty acids induce endoplasmic reticulum stress and apoptosis independently of ceramide in liver cells." in: American journal of physiology. Endocrinology and metabolism, Vol. 291, Issue 2, pp. E275-81, 2006 (PubMed).

Lee, Cherla, Tesh: "Simultaneous induction of apoptotic and survival signaling pathways in macrophage-like THP-1 cells by Shiga toxin 1." in: Infection and immunity, Vol. 75, Issue 3, pp. 1291-302, 2007 (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).

Yuan, Fu, Wang et al.: "Voltage-dependent anion channel 1 is involved in endostatin-induced endothelial cell apoptosis." in: FASEB journal : official publication of the Federation of American Societies for Experimental Biology, Vol. 22, Issue 8, pp. 2809-20, 2008 (PubMed).

Bok, Prikhodko, Green et al.: "Apoptosis in murine norovirus-infected RAW264.7 cells is associated with downregulation of survivin." in: Journal of virology, Vol. 83, Issue 8, pp. 3647-56, 2009 (PubMed).

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