Testing Data
(Jurkat cells lymphocytes) weretreated with 1 muM staurosporinefor 4 hours to induce apoptosis viacaspase activity. Cells were thenstained with either FAM-VAD-OPHI or FLICA FAM-VAD-FMKgreen fluorescent polycaspase inhibitor probes at 10 muM for 1 hour. Cells were washed twiceand read on a BD FACSCantoflowcytometer. Histograms weremade of cell fluorescence intensity output. Staining results with the OPH-based caspase inhibitor (grey solid) are consistent with those achieved using the FMK-based caspase inhibitor (greenoutline). FAM-VAD-OPH caspase inhibitor reagents mirror FAM-VAD-FMKanalogs for in vitro apoptosis detection, and they may be less toxic and more stable.)
FAM-DEVD-OPH in vitro Apoptosis Detection Reagent | FAM-DEVD-OPH detection reagent
FAM-DEVD-OPH in vitro Apoptosis Detection Reagent, Pack of 4 Vials
Synonyms
FAM-DEVD-OPH in vitro Apoptosis; FAM-DEVD-OPH in vitro Apoptosis Detection Reagent; Pack of 4 Vials; FAM-DEVD-OPH detection reagent
Sample Protocol
FAM-OPH Apoptosis Detection Reagents FAM-DEVD-OPH, FAM-VAD-OPH I, and FAM-VAD-OPH II offer a simple yet accurate method to measure caspase activity in cell culture or live animals. Two sample protocols are outlined below.
Target
Apoptosis, Poly Caspases
Excitation / Emission
488-492 nm / 515-535 nm
Method of Analysis
Flow Cytometer, Fluorescence Microscope, Fluorescence Plate Reader
Types of Samples
Cell Culture, Tissue
Preparation and Storage
Store at 2-8 degree C
Testing Data
(Jurkat cells lymphocytes) weretreated with 1 muM staurosporinefor 4 hours to induce apoptosis viacaspase activity. Cells were thenstained with either FAM-VAD-OPHI or FLICA FAM-VAD-FMKgreen fluorescent polycaspase inhibitor probes at 10 muM for 1 hour. Cells were washed twiceand read on a BD FACSCantoflowcytometer. Histograms weremade of cell fluorescence intensity output. Staining results with the OPH-based caspase inhibitor (grey solid) are consistent with those achieved using the FMK-based caspase inhibitor (greenoutline). FAM-VAD-OPH caspase inhibitor reagents mirror FAM-VAD-FMKanalogs for in vitro apoptosis detection, and they may be less toxic and more stable.)
Testing Data
(Jurkat cells lymphocytes) weretreated with 1 muM staurosporinefor 4 hours to induce apoptosis viacaspase activity. Cells were thenstained with either FAM-VAD-OPHI or FLICA FAM-VAD-FMKgreen fluorescent polycaspase inhibitor probes at 10 muM for 1 hour. Cells were washed twiceand read on a BD FACSCantoflowcytometer. Histograms weremade of cell fluorescence intensity output. Staining results with the OPH-based caspase inhibitor (grey solid) are consistent with those achieved using the FMK-based caspase inhibitor (greenoutline). FAM-VAD-OPH caspase inhibitor reagents mirror FAM-VAD-FMKanalogs for in vitro apoptosis detection, and they may be less toxic and more stable.)
Related Product Information for FAM-DEVD-OPH detection reagent
Description: Detect apoptosis in cultured cell settings with the green fluorescent caspase 3/7 inhibitor reagent, FAM-DEVD-OPH. FAM-DEVD-OPH: 5-FAM-Asp-Glu-Val-Asp(OMe)-Oph (2,6-diF) caspase 3/7 inhibitor; Pack of 4 Vials.
Background: Apoptosis Detection with FAM-DEVD-OPH Caspase 3-7 Inhibitor Apoptosis is an evolutionarily conserved process of programmed cell suicide mediated by a cascade of proteolytic enzymes called caspases. Once activated in response to pro-apoptotic signals, caspases cleave protein substrates leading to the eventual disassembly of the cell1. Caspase Inhibitors with FMK Reactive Groups Early research studies using short peptide sequences linked to an aldehyde or fluoromethyl ketone (FMK) reactive group identified a number of peptide sequences preferentially targeted by different caspase enzymes. For example, the sequence Asp-Glu-Val-Asp (DEVD) is cleaved by activated caspase-3 and the sequence Val-Ala-Asp (VAD) demonstrates multiple caspase (activated caspase -1, -2, -3, -4, -5, -6, -7 -8, -9) recognition properties2. When linked to an FMK reactive group, the resulting peptide-FMK molecule will form an irreversible, covalent thioether adduct with the reactive SH-site of caspase enzymes. MyBioSource utilized these findings to develop the often-cited FLICA (FLuorescence Labeled Inhibitors of CAspases) and FLIVO (FLuorescence in vIVO, now discontinued) caspase inhibitor and apoptosis detection reagents, both of which have been widely used for years as simple and reliable methods for screening apoptosis in live cells, tissues, and whole animals. These reagents consist of short peptide sequences with varying caspase specificities, a fluorescent label, such as carboxyfluorescein (FAM), and an FMK reactive moiety. The resulting fluorescent, cell-permeant, non-cytotoxic caspase inhibitor reagents can be added to cell culture media for whole cell apoptosis detection. These labeled caspase inhibitors permeate the cell membrane and form irreversible covalent bonds with activated caspase enzymes inside apoptotic cells, which can then be differentiated from non-apoptotic cells by their retained fluorescence. Caspase Inhibitors with O-phenoxy Reactive Groups MyBioSource has now released a novel set of inhibitor reagents that employ an O-phenoxy (OPH) reactive group in place of an FMK group. In a manner analogous to the FMK class of cysteine reactive compounds, the OPH caspase inhibitors form a stable, covalent thioether adduct with the reactive SH-site of caspase enzymes present in apoptotic cells3. OPH inhibitor compounds have the benefit of being extremely non-cytotoxic, display enhanced stability characteristics compared to the FMK analogs, and have been optimized to provide a high level of performance for in vitro applications3-4. The new FAM-OPH line of green fluorescent in vitro apoptosis detection reagents includes 5-FAM-Val-Ala-Asp(OMe)-2,6-difluorophenoxy-methyl ketone (FAM-VAD-OPH I) poly caspase inhibitor, 5-FAM-Val-Ala-Asp(OMe)-Oph (2,6-diF) (FAM-VAD-OPH II) poly caspase inhibitor, and 5-FAM-Asp-Glu-Val-Asp(OMe)-Oph (2,6-diF) (FAM-DEVD-OPH) caspase 3-7 inhibitor. A red fluorescent poly caspase inhibitor is also available: SR-VAD-OPH.
Background: Apoptosis Detection with FAM-DEVD-OPH Caspase 3-7 Inhibitor Apoptosis is an evolutionarily conserved process of programmed cell suicide mediated by a cascade of proteolytic enzymes called caspases. Once activated in response to pro-apoptotic signals, caspases cleave protein substrates leading to the eventual disassembly of the cell1. Caspase Inhibitors with FMK Reactive Groups Early research studies using short peptide sequences linked to an aldehyde or fluoromethyl ketone (FMK) reactive group identified a number of peptide sequences preferentially targeted by different caspase enzymes. For example, the sequence Asp-Glu-Val-Asp (DEVD) is cleaved by activated caspase-3 and the sequence Val-Ala-Asp (VAD) demonstrates multiple caspase (activated caspase -1, -2, -3, -4, -5, -6, -7 -8, -9) recognition properties2. When linked to an FMK reactive group, the resulting peptide-FMK molecule will form an irreversible, covalent thioether adduct with the reactive SH-site of caspase enzymes. MyBioSource utilized these findings to develop the often-cited FLICA (FLuorescence Labeled Inhibitors of CAspases) and FLIVO (FLuorescence in vIVO, now discontinued) caspase inhibitor and apoptosis detection reagents, both of which have been widely used for years as simple and reliable methods for screening apoptosis in live cells, tissues, and whole animals. These reagents consist of short peptide sequences with varying caspase specificities, a fluorescent label, such as carboxyfluorescein (FAM), and an FMK reactive moiety. The resulting fluorescent, cell-permeant, non-cytotoxic caspase inhibitor reagents can be added to cell culture media for whole cell apoptosis detection. These labeled caspase inhibitors permeate the cell membrane and form irreversible covalent bonds with activated caspase enzymes inside apoptotic cells, which can then be differentiated from non-apoptotic cells by their retained fluorescence. Caspase Inhibitors with O-phenoxy Reactive Groups MyBioSource has now released a novel set of inhibitor reagents that employ an O-phenoxy (OPH) reactive group in place of an FMK group. In a manner analogous to the FMK class of cysteine reactive compounds, the OPH caspase inhibitors form a stable, covalent thioether adduct with the reactive SH-site of caspase enzymes present in apoptotic cells3. OPH inhibitor compounds have the benefit of being extremely non-cytotoxic, display enhanced stability characteristics compared to the FMK analogs, and have been optimized to provide a high level of performance for in vitro applications3-4. The new FAM-OPH line of green fluorescent in vitro apoptosis detection reagents includes 5-FAM-Val-Ala-Asp(OMe)-2,6-difluorophenoxy-methyl ketone (FAM-VAD-OPH I) poly caspase inhibitor, 5-FAM-Val-Ala-Asp(OMe)-Oph (2,6-diF) (FAM-VAD-OPH II) poly caspase inhibitor, and 5-FAM-Asp-Glu-Val-Asp(OMe)-Oph (2,6-diF) (FAM-DEVD-OPH) caspase 3-7 inhibitor. A red fluorescent poly caspase inhibitor is also available: SR-VAD-OPH.
Product Categories/Family for FAM-DEVD-OPH detection reagent
References
Slee, E. A., et. al. Serial killers: ordering caspase activation events in apoptosis. Cell Death Differ. 6:1067-1074 (1999) Thornberry,A., et. al. A combinatorial approach defines specificities of members of the caspase family and granzyme B. J Bio Chem. 272(29):17907-17911 (1997).Chauvier, D., et. al. Broad spectrum caspase inhibitors: from myth to reality? Cell Death Differ. 14:387-391 (2007). Callus, B. A. and Vaux, D. L. Caspase inhibitors:viral, cellular, and chemical. Cell Death Differ. 14:73-78 (2007).
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Product Notes
The FAM-DEVD-OPH (Catalog #AAA258040) is a Detection Reagent and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "FAM-DEVD-OPH in vitro Apoptosis, Detection Reagent" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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