BioScience Catalog Number: - PDF document

Essen BioScience Catalog Number: 4440 Presentation, S torage and S tability The CellPlayer 96 - W ell Kinetic Caspase - 3/7 reagent is provided in one vial (20 µl) of 5 mM DEVD - NucView TM 488 in DMSO capable of performing 100 - 200 tests in a 96 - well plate format . When stored at 4 °C the CellPlayer 96 - W ell Kinetic Caspase - 3/7 reagent is stable for at least six months. Background and Intended Use The CellPlayer 96 - W ell Kinetic Caspase - 3/7 reagent couples the activated caspase - 3/7 recognition motif (DEVD) to NucView TM 488 , a DNA intercalating dye. This reagent , specially formulated by Biotium, Inc. for use in the Essen BioScience IncuCyte ™ FLR or ZOOM system s can be added directly to tissue culture wells using a no wash, mix and read protocol to acquire live cell ima ges of cells undergoing caspase - 3/7 mediated apoptosis. Addition of this reagent is non - perturbing to cell growth and morphology, and enables the kinetic detect ion of caspase - 3/7 activity using the IncuCyte ™ FLR or ZOOM live cell imaging system while never leaving the incubator. When added to tissue culture medium, this inert, non - fluorescent substrate crosses the cell membrane where it is cleaved by activated ca spase - 3/7 resulting in the release of the DNA dye and green fluorescent staining of nuclear DNA. Kinetic activation of caspase - 3/7 can be monitored morphologically using live cell imaging, and quantified using the IncuCyte TM FLR object counting algorithm o r the IncuCyte™ ZOOa basic analyzer. Excitation/Emission Spectra of NucView 488 Fluorescent Product Wavelength (nm) Figure 1. Excitation and emission spectra of enzymatically - cleaved NucVIew 488 caspase - 3 substrate in the presence of excess of dsDNA. CellPlayer™ 96 - W ell Kinetic Caspase - 3/7 Reagent : http://biotium.com/product/product_info/Protocol/psds - 10400.pdf CellPlayer™ 96 - Well Caspas e - 3/7 Reagent 2 Recommended Use We recommend that CellPlayer 96 - W ell Kinetic Caspase - 3/7 reagent be used at a final concentration of 5 µM (1:1000) in growth medium and added directly to cells in culture . When used in an IncuCyte ™ FLR or ZOOM live cell imaging system s , we recommend data collection every 3 hours. Please see the relevant Apoptosis application note published on our webpage at http://www.essenbioscience.com/apoptosis.html for a detailed study using this reagent. MSDS Information MSDS information can be found at: http://biotium.com/wp - content/uploads/2013/11/MSDS - 10402 - 1.pdf For research use only. Not for therapeutic or diagnostic use. Product Label License This product is provided under an Agreement between Biotium and Essen BioScience . The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. (U.S. Patent application No. US60/741,263). The purchase price of this product includes a limited, non - transferable immunity from suit under U.S. Patent application No. US60/741,263 and corresponding patent claims outside of the United States, to use only the purchased amount of the product and components of the product solely for the buyer’s o wn internal research (whether the buyer is an academic or for - profit entity), in a manner consistent with the accompanying product literature. No right to use, sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party, or otherwise use this product or its components or materials made using this product or its components for any other purpose or a Commercial Purpose, is hereby granted expressly, by implication or by estoppel. Commercial Purpose means any activity by a party for consideration and include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service including without limitation, re porting information or data resulting from use of the product or a its components for a fee or other commercial consideration; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or it s components, whether or not such product or its components are resold for use in research. For products that are subject to multiple limited use label licenses, the most restrictive terms apply. This product is for research use only. Diagnostic uses re quire a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265 - 1027. Fax: (510) 265 - 1352. Ema il: btinfo@biotium.com. All names containing the designation ™ are trademarks of Biotium, Inc. 8000 - 0069 - E 00