High Density Lipoprotein HDL cholesterol assay serum 0.9-180 mg/dL liquid stable colorimetric method clin chemistry analyser reagent 1 Wako 4x60ml

High Density Lipoprotein HDL cholesterol Assay Reagent 1, 4x60ml, manufactured by FUJIFILM Wako Pure Chemical Corporation.

Cholesterol is an essential component of cell membranes and required for the synthesis of hormones and bile acids. Cholesterol is transported around the body in lipoprotein complexes which are formed between lipids and apolipoproteins. There are four classes of lipoprotein : high density lipoprotein (HDL), low density lipoprotein (LDL), very low density lipoprotein (VLDL) and chylomicrons.

HDL is responsible for uptake and disposal of cholesterol ('Good Cholesterol') whilst LDL transports cholesterol to around the body where it is deposited in tissues and organs ('Bad Cholesterol'). Excess cholesterol in the blood can be deposited on the walls of blood vessels leading to atherosclerosis and increased risk of heart disease and stroke. HDL Cholesterol is often measured in conjunction with LDL Cholesterol and Total Cholesterol as part of a lipid panel to screen for cardiovascular risk factors and monitor cholesterol reducing therapy.

The Wako HDL Cholesterol assay uses a method of immunoinhibition to determine Cholesterol in serum. The sample is incubated with anti-human beta-lipoprotein antibody (Reagent 1) which binds to lipoproteins other than HDL, forming complexes with LDL, VLDL and chylomicrons which block subsequent enzyme reactions. On addition of Reagent 2, containing cholesterol esterase (CHE) and cholesterol oxidase (CO), only the HDL-Cholesterol is esterified and oxidised resulting in cholestenone, fatty acids and hydrogen peroxide. The hydrogen peroxidase then reacts with F-DAOS [N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxy-4-fluoroaniline, sodium salt] and 4-aminoantipyrine (4-AA) in the presence of peroxidase to give a blue complex. The colour intensity as measured at 600nm can then be used to calculate the HDL-C concentration of the sample.

The Wako HDL-Cholesterol reagents are supplied individually. This product code relates to reagent 1 only, supplied as 4x60ml. HDL-C Reagent 2 (412-72495) and HDL-C Calibrator (416-51095) are sold separately. Both ready-to-use, liquid reagents should be stored at 2-8C and have an open vial stability of 30 days.

The Wako HDL-Cholesterol assay has a measuring range of 0.9-180 mg/dL and should be calibrated with the HDL Calibrator (416-51095). A range of applications for use on clinical chemistry analysers are available - please enquire.

The suggested QCs for this assay are the Wako Control Serums, Normal 410-00104 and Abnormal 416-00204.