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GENTAUR Europe

 GENTAUR Europe BVBA
Voortstraat 49, 1910 Kampenhout BELGIUM
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 GENTAUR BULGARIA
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    Caspase-3 Assay for Apoptosis Detection in Whole, Living Cells 
    Catalog no. 9125 (25-50 tests)

    flica-660-caspase-3-assay-apoptosis-positive-jurkat-cellsApoptosis is an evolutionarily conserved form of cell suicide mediated by a cascade of proteolytic enzymes called caspases. Pro-apoptotic signals activate the enzymatic cascade resulting in the cleavage of protein substrates, leading to the disassembly of the cell. Caspases have been identified in organisms ranging fromC. eleganstoMembers of the mammalian caspase family of cysteinyl aspartate-specific proteases play distinct roles in apoptosis and inflammation.

    Caspases
    Active caspase enzymes exhibit catalytic and substrate specificities comprised of short tetra-peptide amino acid sequences that must contain an aspartate in the P1 position. These preferred tetra-peptide sequences have been used to derive peptides that specifically compete for caspase binding. In addition to the distinctive aspartate cleavage site at P1, the catalytic domains of the caspases typically require four amino acids to the left of the cleavage site with P4 as the prominent specificity-determining residue. Addition of a fluoromethyl ketone (FMK) to the tetrapeptide results in an irreversible linkage and permanent inactivation of the cysteine protease enzyme. Furthermore, conjugation of a fluorescent moiety at the amino terminus yields a probe that allows for the detection of caspase activity.

    FLICA® 660 Caspase-3 Assay: Detection Mechanism

    The far-red FLICA® 660-DEVD-FMK caspase-3 detection probe is comprised of the preferred affinity peptide sequence (DEVD) targeted by activated caspase-3 and caspase-7, a far-red fluorescent 660 dye label, and a fluoromethyl ketone (FMK) reactive moiety. The resulting fluorescent caspase-3 inhibitor probe forms an irreversible, covalent bond with active caspase-3 enzymes, efficiently labeling the target for detection. Due to its cell permeant nature and fluorescence properties, the far-red FLICA caspase-3 detection probe enables whole cell analysis via common fluorescence detection methods.

    To use FLICA Caspase-3 Assay, add the caspase-3 detection probe directly to suspension cell or tissue culture media, incubate, and wash. The cell permeant, far-red FLICA caspase-3 detection probe will efficiently diffuse into cells and irreversibly bind to activated caspase-3 enzymes, thereby retaining the red signal inside caspase-3-positive cells. Cells not bearing active caspase-3 return to a non-fluorescent status after the wash step.

    The FLICA® 660 caspase-3 detection probe has an optimal excitation at 660 nm and optimal emission range from 680-690 nm. As such, it has demonstrated excellent excitation efficiency with a conventional red HeNe laser with a 633 nm excitation, enabling samples to be analyzed with most flow cytometers and fluorescence microscopes equipped with electronic grey scale image capabilities. Cells labeled with the FLICA® caspase-3 detection probe may be read immediately or preserved for 16 hours using the fixative.

    Cat #: 9125 (25-50 tests)

    Price: 250 Euro (without VAT)

     

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    Published in Promos

    flica-660-caspase-3-assay-apoptosis-positive-jurkat-cells

    Catalog Number: 9125 (25-50 tests)

    Quick Overview

    Forget the lysates. Assay for apoptosis via caspase-3 activity in whole, living cells with the FLICA® 660 Caspase-3 Assay Kit.
    - Ex: 660 nm / Em: 680-690 nm
    - Whole cell analysis via flow cytometry or fluorescence microscopy
    - Flexible multiparametric analysis with additional dyes or probes
    - Compatibility with GFP and green autofluorescence
    - Benchtop flow compatible

    This new caspase-3 assay for in vitro apoptosis detection employs a new far-red fluorescent caspase-3 and -7 inhibitor probe, FLICA® 660-DEVD-FMK, to label active caspase-3 and-7 enzymes in living cells or tissue samples. Image the fluorescent signal using fluorescence microscopy or analyze samples for caspase-3 and -7 activity by flow cytometry.

    The cell permeant FLICA caspase-3 detection probe, 660-DEVD-FMK, is comprised of a caspase-3-affinity inhibitor peptide sequence (DEVD) and a fluoromethyl ketone (FMK) moiety that enable an irreversible, covalent binding event with active caspase enzymes. This newest FLICA caspase probe is labeled with a far-red fluorescent 660 dye reporter, enabling caspase-3 detection via common fluorescence detection methods.

    flica-660-caspase-3-assay-apoptosis-positive-jurkat-cells-dic

    Apoptosis detected in induced Jurkat cells by labeling active caspases 3&7 with far-red fluorescent FLICA 660 Caspase 3/7 Assay

     

    Promotional price: 199$

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    Published in Promos