HDL Uptake Assay Kit (Fluorometric)

Only kit on the market to measure HDL uptake
Catalog #: K586 | abID: ab204717

Product Details

abID ab204717
Cat # +Size K586-100
Size 100 assays
Detection Method Fluorescence (Ex/Em 540/575 nm)
Applications Measure HDL uptake in different cell types
Screen, study or characterize stimulators or inhibitors of HDL uptake
Features & Benefits • Only Kit on the market to quantitatively measure HDL uptake.
• Includes unlabeled-HDL for assay validation.
Kit Components • Assay Buffer
• Fluorescently-labeled HDL (5 mg/ml)
• Unlabeled HDL (2x)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


High-density lipoprotein (HDL) consists of a protein shell, containing apolipoproteins A-I and A-II, and a hydrophobic core consisting of cholesterol and triglycerides surrounded by phospholipids and cholesterol esters. HDL transports cholesterol to the liver or steroidogenic organs such as adrenals, ovary, and testes by both direct and indirect pathways. HDL-Cholesterol is removed by HDL receptors such as scavenger receptor BI (SR-BI), which mediate the selective uptake of cholesterol from HDL. Clinically, high levels of HDL has been associated with cardiovascular health. This is due to the ability of HDL particle to transport cholesterol from lipid-laden macrophages of atherosclerotic arteries to the liver for secretion into the bile by a process called as reverse cholesterol transport. BioVision’s HDL Uptake Assay Kit contains a fluorescently-labeled HDL that can be measured when taken up by cells. Unlabeled-HDL is included in the kit for assay validation.

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What is the concentration of BLT1 used for the assay and how long were HepG2 cells incubated with BLT1?
Concentration of BLT1 used was 5 µM with 1-2 hr of pre-incubation. BLT1 was added during treatment with the fluorescently labeled HDL as well.
Can we use frozen samples with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Can we use a different wavelength than recommended for the final analysis?
It is always recommended to use the exact recommended wavelength for the most efficient results. However, most plate readers have flexibility in their band width of detection in increments of +/- 10 nm. Depending on this flexibility range, you can deviate from the recommended wavelengths within limits.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact yopur regional BioVision distributor.
What is the shelf life of this kit?
This kit is good for 12 months from the date of shipment in the unopened form when stored at the appropriate temperature and appropriate conditions. After opening and reconstitution, some of the components in this kit are good for 2 months at -20°C. Please refer to the datasheet for storage information and shelf life of each of the components.
Why are my standard curve values lower than those shown on the datasheet?
There are multiple factors which influence the signals like the incubation times, room temperature, handling etc. In general, to increase the value of the standards, you can increase the incubation time. As long as the standard curve is linear, it should be fine to use, since all of your samples will also be measured under the same conditions on this curve.
How do I normalize my samples against protein concentration
You can use a protein quantitation assay on the supernatants you get from cell/tissue lysates or with any other liquid sample in the assay buffer.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Should I make a standard curve for every expt I do, or is one curve/kit enough?
Yes, I would strongly recommend you to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.
From the data sheet, for determining non-specific labeled HDL, I have to prepare one well as I wrote in an attached excel file, right?
The well were you are treating with competing amount of HDL will actually let you determine the non-specific binding. Any fluorescence that you cannot compete out with extra HDL is actually non-specific binding. So you don't really need to have a dedicated well for that (so the fourth control well is optional)
Can I use FACs?
Yes, it is possible to use cells by FACS.