HPLC Analysis of Thyroglobulin on a SRT® SEC-1000 Gel Filtration Column

HPLC Analysis of Thyroglobulin on a SRT® SEC-1000 Gel Filtration Column

Conditions

column SRT SEC-1000, 30 cm x 4.6 mm I.D., 5 µm particles, 1000 Ã… (Z777061)
column temp. Ambient
mobile phase 150 mM phosphate buffer, pH 7.0
flow rate 0.35 mL/min
sample 1. Thyroglobulin (670kD) 1 mg/mL; 2. Uracil (120D), 0.1 mg/mL
injection 3 µL
detector UV, 214 nm

Description

Analysis Note Thyroglobulin is a very large protein, with a molecular mass of 670 kDa. Thus,a large pore size gel filtration columns is required to prevent partial exclusion of not just the monomer, but also of the dimer and higher order aggregates. This was accomplished using the Sepax SRT SEC-1000 column, which has a nominal pore size of 1000 Angstrom. The nucleic acid base Uracil was added to the sample as a marker to indicate the inclusion volume since a small molecule like Uracil is not restricted from entering any pores in the column.
Categories Analytical Chromatography, Proteins
Featured Industry Life Science and Biopharma
Legal Information SRT is a registered trademark of Sepax Technologies
suitability application for HPLC

Materials

     
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