Ccf2 assay

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August undefined, 2024

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 … 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, …

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

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 … 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 ... holiday inn near austin tx

β-Lactamase: an ideal reporter system for monitoring

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 … Webanalysis protocol. Widely used to assay injection of effector proteins in T3SS and T4SS is an assay that utilizes the enzymatic cleavage of the FRET-reporter cephalosporin CCF2 by injected β-lactamase fu-sions (Charpentier & Oswald, 2004). The CCF2 assay facilitated the analysis of injection kinetics and of intracellular accumulation levels WebMar 9, 2024 · CCF2-AM associated fluorescence for the uncleaved substrate is detected in V7, with cleaved substrate detected in V3. (C) Dot plots showing optimized staining conditions in infected mouse peritoneal cells (PerCs) in vitro. PerCs were extracted and infected overnight in vitro with MHV68 or MHV68.LANA:βlac. holiday inn near banbury

A NanoLuc luciferase-based assay enabling the real-time …

WebSnippet: 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 … 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. hugo\u0027s thief river falls pharmacyWebCCF2-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 … holiday inn near baltimore md

"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 … " - Ccf2 assay

Ccf2 assay

WebSep 13, 2007 · The fluorometric assay is performed with a cephalosporin derivative, CCF2/AM, as the substrate . CCF2/AM contains ester moieties, which facilitate its … 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 …

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WebOct 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 … WebThe GeneBLAzer™ In Vivo Detection Kit measures β-lactamase reporter activity in live mammalian cells, using the β-lactamase substrate CCF2-AM. The GeneBLAzer™ Detection System provides the following …

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 … 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.

WebCCF2/4-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 … 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.

WebSep 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 …

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. hugo\\u0027s thief river falls pharmacyWebCCF2-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 ... hugo\u0027s upholsteryWeb2. 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 holiday inn near banc of california stadiumWebThe 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, hugo\u0027s tree care incWebCCF2-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. hugo\\u0027s tree care incWebNational Center for Biotechnology Information hugo\u0027s thief river falls mnWebThe CCF2-AM and CCF4-AM substrates are essential assay components for the GeneBLAzer™ platform. These substrates are fully compatible with flow cytometry, speeding the time to clone selection. Ratiometric analysis of the blue and green signals reduces well-to-well variation due to differences in cell numbers and substrate loading, … hugo\\u0027s thrifty white drug jamestown nd