| Field | Specification |
|---|---|
| Mfr No | |
| Assay Time | |
| Detection Method | |
| Product Type | |
| Sample Type(s) | Adherent cells |
| Shipping | |
| Species | |
| Storage |
Overview
For quantitatve determination of cholesterol uptake in cells and screening of modulators on cholesterol uptake. The assay uses FL485/535nm for signal readout. Compatible sample input includes Adherent cells. Typical stated assay timing is Assay takes 24-72 hrs, hand-on time 1 hr..
Key elements and design rationale
- Readout format: FL485/535nm supports plate-based signal acquisition and consistent comparison across matched samples.
- Sample compatibility: The stated sample scope includes Adherent cells, which is useful when aligning matrix type with calibration and control design.
- Workflow timing: The listed assay time of Assay takes 24-72 hrs, hand-on time 1 hr. helps frame batch planning, replicate handling, and plate throughput.
- Feature emphasis: Convenient. Treat cells directly in 96-well fluorescent plate.
Additional feature notes highlight Safe. Non-radioactive assay; High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day. Available format information for this listing includes 100 Tests.
Biological background
This product is centered on measurement of cholesterol uptake within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.
More details
CHOLESTEROLis a sterol and lipid present in cell membranes, and is transported in the bloodstream of all animals. It is used to form cell membranes and hormones, and plays important roles in cell signaling processes. Cellular regulation of cholesterol levels is a complex system in which irregularities have been tied to obesity and heart disease. Increased cholesterol uptake has also been linked to highly proliferative cancer cells. Through monitoring cellular cholesterol uptake, one can explore these growing health problems and screen for possible drug treatments. BioAssay Systems cholesterol uptake assay kit is based on cellular uptake of a fluorescently tagged cholesterol probe. The fluorescence intensity measured at λex/em = 485/535 nm is proportional to the amount of cholesterol uptaken by the cells.
Detection method
Fluorescent (FL 485/535 nm).
Procedures and timing
Stated procedure or timing information: Assay takes 24-72 hrs, hand-on time 1 hr.
Research relevance and current trends
- Plate-based quantification and side-by-side group comparison remain central use cases for this assay format.
- The product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.
- The description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.
Common research applications
- Quantify cholesterol uptake in adherent cells by FL485/535 nm readout.
- Compare treatment or phenotype groups using matched adherent cells handling.
- Monitor time-course or pre/post changes in adherent cells across study conditions.
Interpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.
Notes for experimental interpretation
- Matrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.
- Use appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.
The Positive Control isn’t working?
You may need to titrate the Positive Control to determine the optimal concentration for the cell line you are using. Titrate down from the recommended Positive Control concentration of 2.5 µM we suggest in the protocol.
I’m not getting the results I’d expect?
The assay is a cell based assay which intrinsically leaves lofty room for variation in experiments. To account for variation amongst cells, we recommend running each variable in at least duplicate, however, preferably triplicate or greater.
There is no difference in RFU values between wells or RFU values are irregular?
Using high concentration (greater than 1%) serum media for the assay. Serum binds the Fluorescent Tracer making it inaccessible to the cells. Use serum free media or low percentage (less than 1%) serum media.
Not allowing the cells enough time to uptake the cholesterol. The cells need time to uptake the Fluorescent Tracer that is accessible to them.
Wells were not washed. If the wells are not washed after removing the media, some of the Fluorescent Tracer containing media may be left in the well. This can give positive bias and irregular results.
Cells were not aspirated carefully. When removing the media or wash buffer from the wells, be sure to pipette gently to avoid pipetting up cells with the media / wash. The Fluorescent Tracer uptaken by the cells cannot be quantified if the cells are not there.
For laboratories requiring additional technical capacity, we provide scientific support services including assay execution, method guidance, product sourcing, and customization to align the assay with specific experimental objectives. If you need assistance selecting the appropriate kit configuration, adapting the workflow to your application, or identifying related research services, please click Talk to a Scientist, email support@biohippo.com, or review our Research Services; a member of our scientific team will follow up with recommendations tailored to your study.
