Quantification of Plasma Lipoproteins by FPLC

Serum total cholesterol and LDL cholesterol levels were determined using commercial assay kits (#E1005 and #E1018, respectively, from Applygen, Beijing, China). Fast protein liquid chromatography (FPLC) analysis of plasma lipoproteins was performed as described (Guo et al, 2018 (link)). Briefly, pooled plasma aliquots (200 μl from each group) were applied to Tricorn high‐performance Superose S‐6 10/300GL columns and filtered using an Amersham FPLC system (GE Healthcare, Chicago, IL, USA), followed by elution with PBS at a constant flow rate of 0.25 ml/min. Eluted fractions (500 μl per fraction) were assessed for cholesterol concentrations using enzymatic assay (Guo et al, 2018 (link)). FPLC was carried out in an investigator‐blind manner.

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Chen Y., Wu X., Zhang J., Pan G., Wang X., Guo X., Wang J., Cui X., Gao H., Cheng M., Yang J., Zhang C, & Jiang F. (2022). Amino acid starvation‐induced LDLR trafficking accelerates lipoprotein endocytosis and LDL clearance. EMBO Reports, 23(3), e53373.

Publication 2022

E1005

Assay Blind Cholesterol Enzymatic assay Ldl cholesterol Lipoproteins Liquid chromatography Plasma Plasma protein Protein Protein plasma Serum

Corresponding Organization : Chinese Academy of Medical Sciences & Peking Union Medical College

Other organizations : Shandong University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Serum total cholesterol levels
LDL cholesterol levels
Cholesterol concentrations in FPLC fractions

control variables

None explicitly mentioned

controls

Positive control: Not mentioned
Negative control: Not mentioned

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