HPSEC was performed on a BioLogic DuoFlow chromatography
system (BioRad, Hercules, CA). LDH samples (150 μL,
sample concentration), either native or aggregated, after a
number of F/T cycles were loaded onto a prequilibrated
TSKgel SEC column (G3000 SWXL, 7.8 mm×30 cm,
Tosoh Bioscience, Bellefonte, PA), and then eluted with 0.3M
NaCl, 50 mM PBS (pH 7.0). The elution flow rate was 1 ml/
min. LDH aggregation was expressed as the loss of monomer
peak after normalization to the peak area of a native LDH
sample. Three replicates were performed for each LDH
sample to determine the aggregation level after repeated F/T
cycles and the associated standard deviation.
Tidak ada komentar:
Posting Komentar