2024 Ccf2 assay

Ccf2 assay

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August undefined, 2024

WebJan 19, 2012 · The enzymatic conversion of the CCF2/AM substrate was found to be directly related to vector entry, as a neutralizing antibody completely blocked the conversion. ... This rapid assay is based on ... WebThe CCF2 assay facilitated the analysis of injection 95kinetics and of intracellular accumulation levels of effectors (26). It was also successfully used for HTP high content screening of T3SS inhibitors96 (27). Real-time observation of injection was achieved by direct fluorescent labeling of tetracysteine motif-tagged effectors97 (28). However,

Invitrogen™ LiveBLAzer™ FRET-B/G Loading Kit with CCF2-AM

WebFeb 18, 2024 · The CCF2 assay facilitated the analysis of injection kinetics and of intracellular accumulation levels of effectors (Mills, Baruch, Charpentier, Kobi, & Rosenshine, 2008 ). It was also successfully used … 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 … richard wetter obituary

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

WebCCF2-AM and CCF4-AM differ by two carbons in the bridge linking the coumarin moiety to the lactam ring. Both are in the membrane-permeable, esterified forms, and can be used for assays in intact cells. CCF4-AM has better solubility properties (soluble for >24 hours) than CCF2-AM and is thus best suited for screening applications. WebCCF2 is a Fluorescence Resonance Energy Transfer (FRET) substrate which consists of a cephalosporin core linking 7-hydroxycoumarin to fluorescein. This product is to be used as a FRET-based substrate for β … Web2. Add 4 µL 10X tBHQ to Stimulated wells and 4 µL Assay Medium to Unstimulated and Cell-free wells. 3. Incubate the assay plate in a humidified 37°C/5% CO 2 incubator for 15 hours. 5.2.5 Substrate Loading and Incubation This protocol is designed for loading cells with LiveBLAzer™-FRET B/G Substrate (CCF4-AM) or CCF2-AM. If redneck picture

Bioassay Lecture10 Part 3 1 .pdf - Reporter Assays...

CCF2-FA (free acid)

WebWidely used to assay injection of effector proteins in T3SS 93 and T4SS is an assay that utilizes the enzymatic cleavage of the FRET-reporter cephalosporin 94 CCF2 by injected … richard weyand empire seriesWebSnippet: To confirm our results and to obtain further proof for the utilization of DYNA-DEP and DYNA-IND entry routes by IAV, we additionally used an IAV virus-like particle (VLP) direct entry assay [26] . These VLPs contain IAV HA and NA in their envelope and harbor a beta-lactamase reporter protein fused to the influenza matrix protein-1 (BlaM1), which … redneck pictures

"WebDec 15, 2008 · CCF2 fluorescence readings were normalized by dividing fluorescence values measured at 447 nm by those measured at 520 nm, and then normalized to OD 600 of the cells. All data was then normalized to the background fluorescence measured in cells expressing cutinase only. " - Ccf2 assay

Ccf2 assay

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WebMay 16, 2024 · CCF2/4 is composed of two fluorescent dyes, 7-hydroxycoumarin-3-carboxamide and fluorescein, bridged by … WebDec 29, 2024 · The study of fusion of fully infectious HCV particles, HCVcc, was developed later and is based on the fusion of viral particles at the plasma membrane [ 7 ], as previously achieved for other viruses or with liposomes. Again, these assays confirmed the dependency of acid pH and for both E1E2 glycoproteins in cell entry.

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WebAug 20, 2014 · The transfer is monitored by the enzymatic cleavage of CCF2, a fluorescent dye substrate of β-lactamase, loaded into the target cells. Cleavage of the β-lactam ring … Websubstrate (CCF2-AM or CCF4-AM) readily enters the cell. Cleavage by endogenous cytoplasmic esterases rapidly converts CCF2-AM or CCF4-AM into its negatively charged form, CCF2 or CCF4, which is retained in the cytosol. This substrate offers a simple and flexible assay development platform for transcriptional readouts in mammalian cells.

WebCCF2-FA (Free Acid) is a fluorescence resonance energy transfer (FRET) substrate that consists of a cephalosporin core linking B7-hydroxycoumarin to fluorescein. This product is intended for use as a beta-lactamase substrate in a GeneBLAzer™ cell-based assay. GeneBLAzer™ cell-based assays typically utilize the membrane-permeant ester forms ... WebJun 1, 2002 · The in vivo assay uses CCF2/AM (E) (adapted from ref. 6). When no β-lactamase activity is present, excitation of the coumarin at 409 nm leads to FRET to the fluorescein acceptor and emission of ...

WebNov 13, 2009 · This assay also serves as an orthogonal counterscreen to identify epi-absorbance assay artifacts. This biochemical FRET-based assay employs as the beta-lactamase substrate the cephalosporin … WebJan 22, 2003 · CCF2 fluorescence on the BD LSR was collected with the use of 460/50-nm (CCF2 blue fluorescence) and 520/10-nm (CCF2 green fluorescence) bandpass filters …

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WebThis cleavage changes the fluorescence emission spectrum of CCF2-AM from green (520 nm) to blue (447 nm), and thus permits detection of fusion by fluorescence microscopy, … redneck playing cardsWebOct 15, 2004 · This assay involves the use of virions containing β-lactamase-Vpr (BlaM-Vpr) and the loading of target cells with CCF2, a fluorogenic substrate of β-lactamase. Since … richard wexler md los alamitosWebSep 13, 2007 · The fluorometric assay is performed with a cephalosporin derivative, CCF2/AM, as the substrate . CCF2/AM contains ester moieties, which facilitate its … redneckpoint christesWebCCF2/4-AM: Greatest Asset of BLA Reporter Assay After an esterified (acetoxymethylated; AM) form of substrate CCF2/4 (CCF2/4-AM) enters a cell, endogenous esterases convert it to CCF2/4, thereby trapping it inside the cell. Exciting CCF2/4 at 408 nm leads to efficient FRET from the coumarin moiety to the fluorescein derivative and produces green … redneck priorities t shirtsWebCCF2-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 … redneck pool party band richmond vaWebSep 30, 2002 · CCF2 contains a cephalosporin ring linking a 7-hydroxycoumarin to fluorescein. In this configuration, excitation of the coumarin at 409 nm leads to fluorescent resonance energy transfer … richard weyhrich sparta tnWebSep 13, 2007 · The fluorometric assay is performed with a cephalosporin derivative, CCF2/AM, as the substrate ( Fig. 1c ). CCF2/AM contains ester moieties, which facilitate its diffusion across the plasma... redneck plunger ace hardware