One ml of radiolabeled virions was applied over 10 ml of 5 to 40% sucrose gradient in PBS and ultracentrifuged at 27,600 rpm for 18 h at 4°C in a SW44Ti rotor (Beckman). Nine equal fractions were collected from the top and an aliquot of each fraction was removed for density measurement by hand held refractometer (R-5000, Atago, U.S.A). The remainder of each fraction was diluted with 3 volumes of PBS and centrifuged at 39,500 rpm for 16 h at 4°C in a SW55 rotor. The pellet was resuspended in NuPAGE sample buffer and analyzed by SDS-PAGE.
R 5000
Manufactured by Atago
Sourced in United States
The R-5000 is a versatile laboratory equipment designed for precise temperature control and analysis. It features a high-performance heating and cooling system to maintain accurate and stable temperatures within a specified range. The core function of the R-5000 is to provide a controlled environment for various scientific experiments and testing procedures.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using r 5000
1
Sucrose Gradient Purification of Radiolabeled Virions
2
Viral Particle Density Profiling
3
Particle Size Analysis via Laser Diffraction
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Analyses of particle size distribution were carried out using a static laser light diffraction unit (Mastersizer 3000, Malvern Instruments Ltd, Worcestershire, UK). Samples were applied to the instrument with distilled water as dispersion medium at 2,800 rpm until an obscuration rate of 5 % was obtained. The refractive index was determined using a hand-held refractometer (Atago R5000, Atago, Tokyo, Japan).
4
Liquid Refractive Index Measurement System
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Tests were conducted using liquid samples of various known nL (Dodecane (Sigma-Aldrich, anhydrous 99+%, nL = 1.42), 40 M glycerin (Daejung, 99+%) solution (nL = 1.44), 6 M CaCl2 (Kanto, anhydrous 95%) solution (nL = 1.44), and a mixture of 40 M glycerin and 6 M CaCl2 (nL = 1.46) to calibrate the measurement of nL. Mixtures of sucrose (Junsei, guaranteed reagent) and DI water with concentrations varying from 0 to 60 wt%, of which nL vary linearly from 1.33 to 1.44 were used to evaluate the versatility of the proposed measurement system. A commercial hand-held refractometer (ATAGO®, R-5000) having a precision of 0.001 was used to check the accuracy of nL measurement by the proposed system. To avoid the effect of temperature on nL, all experiments were conducted at 25 ± 1 °C. A white light-emitting diode and a diffuser film were used to uniformly illuminate light. To avoid the dispersion effect and illuminate monochromatic light of wavelength 589 nm, we used an orange-colored filter in front of the light source. Images of GSP were captured using a digital single-lens reflex camera (Canon, 650D) and macro lens (Canon, MP-E 65 mm). The captured GSP images were processed using the Image Processing Toolbox™ of MATLAB® R2012a to analyze the brightness profile in the GSP.
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