WebCCF4-AM (LiveBLAzer ™-FRET B/G substrate; Invitrogen) were dissolved separately in acetonitrile, mixed together in a proprietary ratio, and dried down under vacuum. The … WebJul 23, 2024 · Briefly, the lipophilic dye CCF4-AM is absorbed into the cytosol and is cleaved by cytosolic esterases. The resulting dye is retained in the cytosol and produces a green fluorescent signal (525 nm) upon excitation with a violet laser (405 nm) [ 17, 29 ]. When phagosomal permeabilization occurs, the dye becomes accessible to M. marinum.

Mycobacterial ESX-1 secretion system mediates host cell lysis ... - PNAS

WebJan 24, 2024 · CCF4-AM is absorbed into the host cytosol and produces a green fluorescent signal (525 nm) . If the mycobacterial phagosome is permeabilized, then the dye is cleaved by the endogenous Mm cell-surface–associated β-lactamase BlaC, which is otherwise inaccessible to the dye. Cleavage causes a loss of FRET and an increase in blue … WebCCF2-FA is de-esterified and used in cell lysate applications, bypassing loading across the cell membrane and de-esterification steps. Cell lysates are the preferred method for applications using cells that contain a cell wall. CCF2-FA can also be used as a control to acquire the excitation and emission spectra for CCF2-AM and CCF4-AM. inflict debuffs on enemies at least once

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WebCCF4-AM is a FRET probe consisting of two fluorophores attached by a β-lactam linkage. ... Cbp1, a fungal virulence factor under positive selection, forms an effector complex that drives ... WebJun 1, 2015 · Cells were then loaded with CCF4-AM, fixed, and intracellularly stained with antibodies against IL-12p40. Plots show cytokine production at 16 h postinfection (PI). Line graph shows Δ flaA -infected macrophages where total = the total frequency of cells positive for the given cytokine. Web1. ™Prepare Solution A: 1 mM LiveBLAzer -FRET B/G Substrate (CCF4-AM, MW = 1096) stock solution in dry DMSO. Store the aliquots of the stock solution at -20°C until use. 2. ™Prepare 6X LiveBLAzer -FRET B/G (CCF4-AM) Substrate Mixture: Add 6 µl of Solution A to 60 µl of Solution B and vortex. inflict corporal inj on spouse/cohab 273.5 pc