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Pigments were eluted using a lineal gradient of 100%% A to 100%% B within the first 12 min, followed by isocratic elution with 100%% B during the next 6 min. Absorbance was monitored at 445 nm.
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The soluble protein was separated from the cell debris by centrifugation at 12,000 × g for 15 min at 4°C, then loaded onto a 5 mL GSTrap FF column equilibrated with buffer A and eluted with a lineal gradient of 50 mM Tris-Cl and 10 mM reduced glutathione at pH 8.1 (buffer B).
Each gradient was fractionated into 40 fractions using a Density Gradient Fractionation System (Brandel, Gaithershurg, MD).
Interaction chromatography is generally performed using a solvent gradient or a temperature gradient.
The gradients were eluted using a Brandel Gradient Fractionator and prepared for qMS and EM analysis.
Gradients were eluted using a gradient fractionator (Brandel) and monitored with a UA-5 detector (ISCO).
A 10 50% sucrose gradient in buffer D was prepared using a gradient mixer.
using a standard sizing gradient.
The data were analyzed using a general lineal model with the Statistical Parametric Mapping (SPM) software package [46].
Samples were analyzed using a gradient described previously [6].
We evaluated predictive factors of increased IL-6, CRP levels using a multivariate lineal regression model on log-transformed data.
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