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-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.

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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 … 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 … the bridges of madison county film location

Using the β-lactamase protein-fragment …

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, 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 … 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 … tart shop richmond