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Cholesterol Efflux Fluorometric Assay Kit (cell-based)

based on 1 citations in multiple journalsCholesterol Efflux Fluorometric Assay Kit (cell-based)14.1 4
Only Assay on the market to measure cholesterol efflux, Non-radioactive, High-Throughput
Catalog #: K582

In stock

$845.00

Product Details

Cat # +Size K582-100
Size 100 assays
Detection Method Fluorescence (Ex/Em = 482/515 nm).
Applications Screen serum samples or lipoproteins (isolated or recombinant) for cholesterol efflux.
Screen small molecules for their effect on cholesterol efflux (a valuable tool for drug discovery program).
Features & Benefits • Simple & Rapid Protocol
• Convenient: Non-Radioactive, no special handling or disposal required
• High-Throughput
• Accurate: reproducible results with low intra & inter assay variability
Kit Components • Labeling Reagent
• Equilibration Buffer
• Reagent A
• Reagent B
• Cell Lysis Buffer
• Positive Control
• Serum Trea™ent Reagent
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Cholesterol efflux from the peripheral tissues and cells in atherosclerotic plaque is an initial and critical step in Reverse Cholesterol Transport (RCT). RCT is the process by which extrahepatic cells, including macrophage-derived foam cells in arterial atherosclerotic plaque, transport excessive cholesterol back to the liver for bile acid synthesis and excretion, thus lowering the peripheral lipid burden. A negative correlation has been established between the in vitro efflux of cholesterol from macrophages and atherosclerosis. BioVision’s Cholesterol Efflux Assay is a high-throughput screening assay for measuring cholesterol efflux in cells using fluorescently-labeled cholesterol. This assay provides a safe, sensitive, and reproducible method for measuring cholesterol efflux.


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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.
What is the purpose of the serum treatment reagent?
The serum treatment reagent gets rid of the proteins and complement from the serum.
What exactly is the positive control in this assay? Or what is the use of the positive control reagent supplied?
The positive control is a chemical that induces cholesterol efflux. This is to help validate that the assay works in the particlar cells being used.
What are the functions of Reagent A and Reagent B?
The function of reagent A is to prevent the degradation of cholesterol in the cell and that of reagent B is to provide the right conditions for labeling.
Yang LY et al., (2017) Angiopoietin‐Like Protein 4 Is a High‐Density Lipoprotein (HDL) Component for HDL Metabolism and Function in Nondiabetic Participants and Type‐2 Diabetic Patients, J Am Heart Assoc, 2017, 6(6): epub
For more citations of this product click here