| Field | Specification |
|---|---|
| Assay Time | |
| Detection Method | |
| Product Type | |
| Sample Type(s) | Cell culture media |
| Shipping | |
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| Storage |
Overview
For quantitative assay of adipolysis and evaluation of drug effects on adipolysis. The assay uses OD570nm, or FL530/585nm for signal readout. Compatible sample input includes Cell culture media. Typical stated assay timing is 20 min.
Key elements and design rationale
- Readout format: OD570nm, or FL530/585nm supports plate-based signal acquisition and consistent comparison across matched samples.
- Sample compatibility: The stated sample scope includes Cell culture media, which is useful when aligning matrix type with calibration and control design.
- Analytical range context: The supplied specifications include a stated detection limit of 0.92 µg/mL for interpreting low-signal samples.
- Feature emphasis: Sensitive and accurate. Use as little as 10 µL samples. Linear detection range in 96-well plate: 0.92 to 100 µg/mL (10 to 1000 µM) glycerol for colorimetric assays and 0.2 to 5 µg/mL for fluorimetric assays.
Additional feature notes highlight Rapid and convenient. The procedure involves addition of a single working reagent and incubation for 20 min at room temperature; Robust and amenable to HTS assays. Potential interference by testing drugs is greatly reduced at 570nm. Compatible with culture media containing phenol red. Assays can be performed in 96 or 384-well plates. Available format information for this listing includes 200 Tests.
Biological background
This product is centered on measurement of adipolysis 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
Obesity is a chronic condition that develops from the storage of excess energy in the form of adipose tissue. The resulting adiposity presents a high-risk factor for diseases such as type 2 diabetes, cardiovascular diseases, and cancer. ADIPOLYSIS or lipolysis is a highly regulated process in fat metabolism, in which triglycerides are broken down into glycerol and free fatty acids. Rapid, robust and accurate procedures for adipolysis quantification in cell culture are very useful in research and drug discovery. BioAssay Systems adipolysis assay kit directly measures glycerol released during adipolysis. This homogeneous assay uses a single Working Reagent that combines glycerol kinase, glycerol phosphate oxidase and color reactions in one step. The color intensity of the reaction product at 570nm is directly proportional to glycerol concentration in the sample.
Detection method
Colorimetric (OD 570 nm) or Fluorescent (FL 530/585 nm).
Detection limit and analytical sensitivity
Reported detection limit: 0.92 µg/mL.
Procedures and timing
Stated procedure or timing information: 20 min.
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 adipolysis in cell culture media by OD570 nm, or FL530/585 nm readout.
- Compare treatment or phenotype groups using matched cell culture media handling.
- Monitor time-course or pre/post changes in cell culture media 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.
I collected the medium from the culture wells. Do I need to now centifuge the medium and collect the supernatant or do I use the medium itself for the assay?
You can assay the cell culture medium directly. However, if it contains cellular debris you should first centrifuge it and then use the clear supernatant in the assay.
What is the principle of the assay?
In the process of adipolysis triglycerides are hydrolyzed by the cellular lipases into glycerol and free fatty acids. The assay measures glycerol, the end product of adipolysis, in a coupled three step enzyme reaction:
1) Glycerol kinase converts glycerol and ATP to glycerol phosphate and ADP.
2) Glycerol phosphate oxidase converts glycerol phosphate and O2 to glycerone phosphate and H2O2
2
2
2
3) HRP converts ADHP and H2O2 to resorufin and water.
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.
Nicotinamide riboside enhances in vitro beta-adrenergic brown adipose tissue activity in humans
Nascimento, EBM et al (2021). Nicotinamide riboside enhances in vitro beta-adrenergic brown adipose tissue activity in humans. The Journal of Clinical Endocrinology and Metabolism. Assay: Adipolysis in human adipocytes.
Presence of vasculature results in faster insulin response in adipocytes in vascularized adipose tissue model
Huttala, Outi, et al (2019). Presence of vasculature results in faster insulin response in adipocytes in vascularized adipose tissue model. ALTEX-Alternatives to animal experimentation. Assay: Adipolysis in human adipose tissue.
Short-chain fatty acids differentially affect intracellular lipolysis in a human white adipocyte model
Jocken, Johan WE, et al (2018). Short-chain fatty acids differentially affect intracellular lipolysis in a human white adipocyte model. Frontiers in endocrinology 8: 372. Assay: Adipolysis in human stem cell.
Development of a three-dimensional adipose tissue model for studying embryonic exposures to obesogenic chemicals
Wang, Rebecca Y., et al (2017). Development of a three-dimensional adipose tissue model for studying embryonic exposures to obesogenic chemicals. Annals of biomedical engineering 45.7: 1807-1818. Assay: Adipolysis in human tissues.
Reconstitution of UCP1 using CRISPR/Cas9 in the white adipose tissue of pigs decreases fat deposition and improves thermogenic capacity
Zheng, Qiantao, et al (2017). Reconstitution of UCP1 using CRISPR/Cas9 in the white adipose tissue of pigs decreases fat deposition and improves thermogenic capacity. Proceedings of the National Academy of Sciences 114.45: E9474-E9482. Assay: Adipolysis in pig tissues.
Differentiation of human adipose stromal cells in vitro into insulin-sensitive adipocytes
Huttala, Outi, et al (2016). Differentiation of human adipose stromal cells in vitro into insulin-sensitive adipocytes. Cell and tissue research 366.1: 63-74. Assay: Adipolysis in human cells.
An apolipoprotein B100 mimotope prevents obesity in mice
Kim, Hyo Joon, et al (2016). An apolipoprotein B100 mimotope prevents obesity in mice. Clinical Science 130.2: 105-116. Assay: Adipolysis in mouse cells.
Rutin ameliorates obesity through brown fat activation
Yuan, Xiaoxue, et al (2016). Rutin ameliorates obesity through brown fat activation. The FASEB Journal 31.1: 333-345. Assay: Adipolysis in mouse cells.
Effect of Nelumbo nucifera Petal Extracts on Lipase, Adipogenesis, Adipolysis, and Central Receptors of Obesity
Chandrasekaran V., Amit A. et al (2013). Effect of Nelumbo nucifera Petal Extracts on Lipase, Adipogenesis, Adipolysis, and Central Receptors of Obesity. Evidence-Based Complementary and Alternative Medicine. 2013: 10.1155/2013/145925 Assay: Adipolysis in cell lines.
Neuropeptide Y potentiates beta-adrenergic stimulation of lipolysis in 3T3-L1 adipocytes
Li R et al (2012). Neuropeptide Y potentiates beta-adrenergic stimulation of lipolysis in 3T3-L1 adipocytes. Regul Pept. 178(1-3):16-20. Assay: Adipolysis in mouse adipocytes.
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