FAM-DEVD-OPH detection reagent product blog
Tags: Detection Reagent; FAM-DEVD-OPH in vitro Apoptosis; FAM-DEVD-OPH detection reagent; FAM-DEVD-OPH;
The FAM-DEVD-OPH n/a (Catalog #MBS258040) is a Detection Reagent and is intended for research purposes only. The product is available for immediate purchase.To buy or view more detailed product information and pricing, please click on the technical datasheet page below:
Please refer to the product datasheet for known applications of a given detection reagent. We\'ve tested the FAM-DEVD-OPH in vitro Apoptosis Detection Reagent, Pack of 4 Vials with the following immunoassay(s):
Testing Data (Jurkat cells lymphocytes) were
treated with 1 muM staurosporine
for 4 hours to induce apoptosis via
caspase activity. Cells were then
stained with either FAM-VAD-OPH
I
or FLICA FAM-VAD-FMK
green fluorescent poly
caspase inhibitor probes at 10 muM
for 1 hour. Cells were washed twice
and read on a BD FACSCanto
flowcytometer. Histograms were
made 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 (green
outline). FAM-VAD-OPH caspase inhibitor reagents mirror FAM-VAD-FMK
analogs for in vitro apoptosis detection, and they may be less toxic and mor
e stable.
)
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.