WebApr 30, 2024 · FITC shows a pH-dependent variation in fluorescence when analyzed at maximum emission wavelength and no variation when analyzing at the isosbestic point, thereby the ratio can be used to determine the lysosomal pH. A standard curve is obtained by equilibrating intralysosomal pH with extracellular pH using the ionophore nigericin. WebA = Slope of the FITC standard curve (ΔRFU/pmol), C = µl of Sample used in the assay, P = Protein concentration in the lysate (mg/ml). 1000 = Conversion Factor (1000 µl ≡ 1 ml) Unit Definition: 1 U is the amount of Collagenase required to cleave the Collagenase Substrate and release 1 pmol of Fluorescein per min under the conditions of the ...
Albumin, Fluorescein isothiocyanate Conjugate bovine (A9771) - Product
WebPlot the FITC Standard Curve. Apply the ∆RFU to the FITC Standard Curve to get B nmol of FITC (amount of unquenched FITC generated between T 1 and T 2 Protease Activity = 𝑩 (𝐓 −𝐓 )× 𝐕 × Sample Dilution Factor = nmol/min/ml = mU/ml Where: B is the FITCamount from Standard Curve (in nmol). 1 is the time of the first reading (R 1 WebBriefly, FITC-inulin (5% in 0.85% NaCl) is dialyzed for 24 h against 0.85% NaCl which results in a ~2% solution and serves to establish the standard curve. The dialyzed FITC-inulin solution is sterile filtered and injected into the retro- orbital plexus (2 μl/g/body weight) during brief isoflurane anesthesia. childcare grant scheme for providers
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Webof FITC-dextran permeability, the Plasma to PBS ratio must be consistent throughout the standard curve, as well as with the dilution factor of the sample. Use normal mouse serum diluted in PBS for the blank. 6. Transfer 50-100 ul of supernatant to a 96-well plate and read on a fluorescent plate reader. WebA standard curve made with FITC-Dextran diluted in PBS References Aoki S, Y.K., Yokoyama T, Nonogaki T, Iwasaki S, Mitsui T, Niwa S., Role of enteric bacteria in the … http://fmrc.pulmcc.washington.edu/DOCUMENTS/FMRC3.pdf child care grants