產(chǎn)品詳情

  • 產(chǎn)品名稱：Caspase-3SubstrateDEVD-pNA

  • 產(chǎn)品型號：
  • 產(chǎn)品廠商：Biovision
  • 產(chǎn)品文檔：
你添加了1件商品 查看購物車
簡單介紹：
Caspase-3SubstrateDEVD-pNA
詳情介紹：
Sequence Ac-Asp-Glu-Val-Asp-pNA
Purity > 98?% by HPLC
Chemical Name Ac-DEVD-pNA, Caspase-3 Substrate, Colorimetric
Formula C??H??N?O??
Permeability Not-permeable
Molecular Weight 638.58 g/mol
Comment

Ready-to-use colorimetric substrate for CPP32/caspase-3 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 spectrophotometric detection of free pNA (l = 400 nm) after cleavage from the peptide substrate DEVD-pNA, using a spectrophotometer or multi-well plate reader. The ready-to-use caspase substrate provides an economic alternative for researchers who perform large amounts of caspase assays.
Note: Background reading from cell lysates and buffers should be subtracted from the readings of both induced and the uninduced samples before calculating fold increase in caspase activity.
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.
3. Resuspend cells in 50 μL of chilled Cell Lysis Buffer and incubate cells on ice for 10 minutes.
4. Centrifuge for 1 min in a microcentrifuge (10,000 x g).
5. Transfer supernatant (cytosolic extract) to a fresh tube and put on ice.
6. Assay protein concentration.
7. Dilute 50-200 μg protein to 50 μL Cell Lysis Buffer for each assay.
8. Add 50 μL of 2X Reaction Buffer containing 10 mM DTT to each sample.
9. Add 5 μL of the 4 mM of DEVD- p NA (200 μM final conc.) and incubate at 37 °C for 1-2 hour.
9. Read samples at 400- or 405-nm in a microtiter plate reader, or spectrophotometer using a 100- μL micro quartz cuvet (Sigma), or dilute sample to 1 mL with Dilution Buffer and using regular cuvet (note: Dilution of the samples proportionally decreases the reading). You may also perform the entire assay directly in a 96-well plate. Fold-increase in caspase activity can be determined by comparing these results with the level of the uninduced control. Note: Background reading from cell lysates and buffers should be subtracted from the readings of both induced and the uninduced samples before calculating fold increase in caspase activity.
Restrictions For Research Use only
Format Liquid
Handling Advice Protect from light and moisture
Storage -20 °C
Expiry Date 6-12 months
Product cited in: Lin, Chien, Tseng, Chen, Chang, Lin: "Furano-1,2-Naphthoquinone Inhibits Src and PI3K/Akt Signaling Pathways in Ca9-22 Human Oral Squamous Carcinoma Cells." in: Integrative cancer therapies, Vol. 13, Issue 3, pp. NP18-NP28, 2014 (PubMed).

Feinman, Deitch, Watkins, Abungu, Colorado, Kannan, Sheth, Caputo, Lu, Ramanathan, Attan, Badami, Doucet, Barlos, Bosch-Marce, Semenza, Xu: "HIF-1 mediates pathogenic inflammatory responses to intestinal ischemia-reperfusion injury." in: American journal of physiology. Gastrointestinal and liver physiology, Vol. 299, Issue 4, pp. G833-43, 2010 (PubMed).

Whitlock, Valbuena, Popov, Judy, Estes, Torres: "Burkholderia mallei cellular interactions in a respiratory cell model." in: Journal of medical microbiology, Vol. 58, Issue Pt 5, pp. 554-62, 2009 (PubMed).

Yuan, Fu, Wang, Shi, Huang, Song, Li, Song, Luo: "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).

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).

Goga, Chiba, Shimizu, Mitani: "Compressive force induces osteoblast apoptosis via caspase-8." in: Journal of dental research, Vol. 85, Issue 3, pp. 240-4, 2006 (PubMed).

Wolf, Brüss, H?nisch, G?thert, von Kügelgen, Molderings: "Molecular basis for the antiproliferative effect of agmatine in tumor cells of colonic, hepatic, and neuronal origin." in: Molecular pharmacology, Vol. 71, Issue 1, pp. 276-83, 2006 (PubMed).

Li, Prabhakaran, Mills, Borowitz, Isom: "Enhancement of cyanide-induced mitochondrial dysfunction and cortical cell necrosis by uncoupling protein-2." in: Toxicological sciences : an official journal of the Society of Toxicology, Vol. 86, Issue 1, pp. 116-24, 2005 (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).

Caporali, Imai, Altucci, Cancemi, Caristi, Cicatiello, Matarese, Penta, Sarkar, Bresciani, Weisz: "Distinct signaling pathways mediate stimulation of cell cycle progression and prevention of apoptotic cell death by estrogen in rat pituitary tumor PR1 cells." in: Molecular biology of the cell, Vol. 14, Issue 12, pp. 5051-9, 2003 (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).

Shain, Landowski, Dalton et al.: "Adhesion-mediated intracellular redistribution of c-Fas-associated death domain-like IL-1-converting enzyme-like inhibitory protein-long confers resistance to CD95-induced apoptosis in hematopoietic ..." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 168, Issue 5, pp. 2544-53, 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).

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).

Ding, Lin, McGill, Juo, Zhu, Blenis, Yuan, Fisher: "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).

Datta, Oki, Endo, Biedermann, Ren, Kufe: "XIAP regulates DNA damage-induced apoptosis downstream of caspase-9 cleavage." in: The Journal of biological chemistry, Vol. 275, Issue 41, pp. 31733-8, 2000 (PubMed).

Doostzadeh-Cizeron, Yin, Goodrich: "Apoptosis induced by the nuclear death domain protein p84N5 is associated with caspase-6 and NF-kappa B activation." in: The Journal of biological chemistry, Vol. 275, Issue 33, pp. 25336-41, 2000 (PubMed).

Fujii, García-Bermejo, Bernabó, Caama?o, Ohba, Kuroki, Li, Yuspa, Kazanietz: "Involvement of protein kinase C delta (PKCdelta) in phorbol ester-induced apoptosis in LNCaP prostate cancer cells. Lack of proteolytic cleavage of PKCdelta." in: The Journal of biological chemistry, Vol. 275, Issue 11, pp. 7574-82, 2000 (PubMed).

Gómez-Angelats, Bortner, Cidlowski: "Protein kinase C (PKC) inhibits fas receptor-induced apoptosis through modulation of the loss of K+ and cell shrinkage. A role for PKC upstream of caspases." in: The Journal of biological chemistry, Vol. 275, Issue 26, pp. 19609-19, 2000 (PubMed).

Kown, Van der Steenhoven, Blankenberg, Hoyt, Berry, Tait, Strauss, Robbins: "Zinc-mediated reduction of apoptosis in cardiac allografts." in: Circulation, Vol. 102, Issue 19 Suppl 3, pp. III228-32, 2000 (PubMed).