Overview

Product name
HDL and LDL/VLDL Cholesterol Assay Kit

See all HDL LDL/VLDL products (2) ...


Product overview
The HDL and LDL/VLDL Cholesterol Assay Kit provides a simple method for convenient separation of HDL (high-density lipoprotein) from LDL (low-density lipoprotein) and VLDL (very low-density lipoprotein) in serum sample, then HDL and LDL/VLDL cholesterol levels can easily be quantified separately, using cholesterol oxidase mediated reactions. Cholesterol esterase hydrolyzes cholesteryl ester into free cholesterol, which can then be oxidized by cholesterol oxidase to yield H2O2. The H2O2 interacts with a sensitive cholesterol probe to produce resorufin, which can be detected with a micro-plate reader at OD = 570 nm or fluorescence micro-plate reader at Ex/Em = 538/587 nm. The assay can also detect cholesterol ester and free cholesterol separately in these two fractions by adding or not adding cholesterol esterase into the reactions.


Notes
The kit can be optimized for cell samples in the following manner:



10-50 mg of tissue must be homogenized well in the assay buffer using a dounce homogenizer. This must be centrifuged at full speed for 5 minutes. The supernantant obtained can be used as follows

” Mix 100 ul of 2X Precipitation Buffer with 100 ul of (cell supernatant)/ serum sample in microcentrifuge tubes. Incubate 10 min at room temperature, centrifuge at 2000 x g (5000 rpm on bench-top microcentrifuge) for 10 min. Transfer the supernatant into new labeled tubes. This is the HDL fraction. The precipitates are the LDL/VLDL fraction. If you want to measure the LDL/VLDL level, the precipitate should be spun again, and the trace amount of HDL supernatant carefully removed. Resuspend the precipitate in 200 ul PBS (not provided). This is the LDL/VLDL fraction.”


Tested applications
Functional Studiesmore details

Properties

Storage instructions
Store at -20°C, Stable for 6 months at -20°C



    

        

            
Components
            
Identifier
            
100 tests
        
    
    

        

            
2X LDL/VLDL Precipitation Buffer
            
NW
            
1 x 10ml
        
        

            
Cholesterol Assay Buffer
            
WM,
            
1 x 25ml
        
        

            
Cholesterol Esterase (Lyophilized)
            
Blue
            
1 vial
        
        

            
Cholesterol Probe (in DMSO, anhydrous)
            
Red
            
1 x 200µl
        
        

            
Cholesterol Standard (2 ug/ul)
            
Yellow
            
1 x 100µl
        
        

            
Enzyme Mix (Lyophilized)
            
Green
            
1 vial
        
    


http://www.abcam.com/HDL-and-LDL-VLDL-Cholesterol-Assay-Kit-ab65390.html