LiveBLAzer™ FRET-B/G Loading Kit with CCF2-AM, 5 mg

LiveBLAzer™ FRET-B/G Loading Kit with CCF2-AM, 5 mg

GeneBLAzer™ cell-based assays utilize the membrane-permeant ester forms (CCF2-AM and CCF4-AM) of the negatively charged fluorescent beta-lactamase substrates, CCF2 and CCF4. These lipophilic esters readily enter the cell, where cleavage by endogeneous cytoplasmic esterases rapidly converts them into their negatively charged forms, thereby trapping them in the cytosol. Detection of GeneBLAzer™ assays is FRET-based. Each substrate is labeled with two fluorophores that form an efficient FRET pair. In the absence of beta-lactamase activity, exciting the coumarin at 409 nm in the intact CCF2 molecule results in FRET to the fluorescein, which emits a green fluorescence signal at 518 nm (Figure 1). In the presence of beta-lactamase activity, however, cleavage of CCF2 spatially separates the two dyes and disrupts FRET, so that exciting the coumarin at 409 nm now produces a blue fluorescence signal at 447 nm. This blue signal can be readily observed under a microscope and can also be detected as an increase in the blue channel readout on fluorescent microplate readers

Features

• For Use With (Equipment):Flow Cytometer, Microplate Reader

Contents

• CCF2-AM or CCF4-AM
• DMSO (for Solution A)
• Solution B
• Solution C