Abstract
When total protein in very low density lipoprotein samples is measured by the method of Lowry et al. (1951. J. Biol. Chem 193:265-275), turbidity remains in the final color reaction. Addition of 0.1 ml of 2.5% (v/v) solution of Triton X-100 removes this turbidity immediately and effectively. This simple modification is faster, less cumbersome, and more economical than removing turbidity with ethyl ether or chloroform. Addition of 0.1 ml of 2.5% (w/v) sodium dodecyl sulfate to the final reaction mixture is also effective in removing turbidity and can be used as an alternative method.
Highlights
The single wash chloroform method gave a mean concentration that was 19.8%higher than the results obtained by the Triton X- 100 method
The concentrations obtained by the single chloroform wash method were consistently higher than results obtained by the Triton X100 method
An exception was the chloroform method where a greater degree of scatter was observed for the same set of samples measured by the Triton X- 100method
Summary
Venous blood was drawn from normal subjects and patients being managed for hypertriglyceridemia at the Lipid Clinic. Blood was drawn in tubes containing the disodium salt of ethylenediaminetetraacetic acid (EDTA) to yield a final concentration of 1.5 mg/ml blood. Following centrifugation at 4°C to obtain plasma, VLDL was isolated using a 40.3 Beckman rotor ( 1 1 ). The supernate from the first ultracentrifugation was layered with EDTA-saline and spun to obtain ‘washed’VLDL. Triton X-100 was purchased from Packard Instrument Company, Inc., Downers Grove, IL. Ethyl ether was purchased from Burdick and Jackson Laboratories, Inc., Muskegon, MI. Chloroform was obtained from Matheson, Coleman, and Bell, Norwood, OH. Sodium dodecyl sulfate was purchased from Canalco, Inc., Rockville, MD. Crystalline bovine serum albumin (BSA) was obtained from Sigma Chemical Co., St. Louis, MO
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