Blood samples were collected from mice (8–12 weeks old) via the tail vein and centrifuged at 3000 rpm for 10 minutes to isolate serum for the measurement of serum levels of total cholesterol (TC), triglyceride, high-density lipoprotein cholesterol (HDL-C), free fatty acid (FFA), ketone bodies, as well as AST (aspartate aminotransferase), and ALT (alanine aminotransferase) activity, using their specific colorimetric kits according to the manufacturer’s instructions. TC, triglycerides, and HDL-C kits were from BioSino Biotechnology and Science, Inc, AST and ALT kits from Nanjing Jiancheng Bioengineering Institute, and ketone bodies from Solarbio (China). Fast protein liquid chromatography was performed on an AKTA system (GE, ÄKTA pure 25 L1).
Kta pure 25 l1
Manufactured by GE Healthcare
The ÄKTA pure 25 L1 is a chromatography system designed for purification of biomolecules. It is a modular system capable of performing a variety of chromatographic techniques such as ion exchange, affinity, and size exclusion chromatography. The system is equipped with a variety of detectors and can be automated for unattended operation.
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2 protocols using kta pure 25 l1
1
Comprehensive Lipid and Enzyme Profiling
2
Determining Dimeric State of Purified Proteins
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To determine their dimeric state in solution, an equal amount of purified apo-CPB, CPB CP, or CPB CP_v1 (see the purification method below) was loaded onto Superdex 75 Increase 10/300 GL (GE Healthcare) connected to ÄKTA Pure 25 L1 (GE Healthcare). Samples were eluted in a buffer [20 mM Tris-HCl (pH 7.5), 500 mM NaCl, 5% (v/v) glycerol, and 0.5 mM TCEP] at the flow rate of 0.5 mL/min for 60 min. The data were collected and analyzed by Unicorn 7 software (GE Healthcare). An equal amount of BSA (Sigma-Aldrich) was analyzed under the same conditions for comparison.
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