Product nameCholesterol Assay Kit - HDL and LDL/VLDL
See all HDL LDL/VLDL kits
Sample typeUrine, Serum, Plasma, Other biological fluids, Tissue Lysate
Assay time1h 00m
Cholesterol Assay Kit ab65390 uses a simple method to quantify total cholesterol, free cholesterol, and cholesterol esters in mammalian samples. It also includes an easy method to separate HDL and LDL / VLDL cholesterol.
In the cholesterol assay protocol, cholesterol oxidase acts on free cholesterol to produce a chemical which reacts with a probe to generate color (570 nm) and fluorescence (Ex/Em = 538/587 nm). Cholesterol esterase is used to hydrolyze cholesteryl ester into free cholesterol.
If cholesterol esterase is included in the assay, total cholesterol is measured. If it is not included, free cholesterol is measured. The amount of cholesterol ester can be calculated by subtracting free cholesterol from total cholesterol.
Cholesterol assay protocol summary:
- use complete sample for cholesterol measurement; or to separate HDL and LDL/VLDL, mix sample with precipitation buffer, incubate for 10 min, centrifuge for 10 min at 2,000 g, keep HDL fraction supernatant, repeat centrifugation and resuspend LDL/VLDL fraction pellet
- for assay, add samples and standards to wells
- add total cholesterol reaction mix (with esterase) or free cholesterol reaction mix (without esterase) and incubate for 60 min at 37ºC
- analyze with microplate reader
If you need to purchase additional HDL precipitation buffer, please see ab105138.
Review our Metabolism Assay Guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.
Storage instructionsStore at -20°C. Please refer to protocols.
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
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Cholesterol Metabolism
RelevanceHDL (high-density-lipoprotein), LDL (low-density lipoprotein) and VLDL (very low-density lipoprotein) are lipoproteins that act as carrier proteins for cholesterol. The regulation of HDL-cholesterol and LDL-cholesterol plays a central role in various disease development. It is well known that low levels of HDL and high level of LDL are associated with an increased risk of cardiovascular events.
- High density lipoprotein
- Low density lipoprotein
Wild type (WT) or farnesoid X receptor knock-out (FRX-KO) mice were treated with either vehicle (Veh) or a high fat-containing diet (HFD) for 16 weeks. Serum HDL (top) and LDL/VLDL cholesterol (bottom) from 5-7 mice/group was measured using ab65390 following protocol instructions. An asterisk (*) means P<0.05 between WT and FXR-KO vehicle group. A pound (#) indicates P<0.05 between Wt and FXR-KO-HFD group. Image obtained from Li G et al., PLoS One, 2012; 7(4): e35895
Rat serum samples were processed according to the protocol. The quantity of total (dilution range 1:10-1:100), HDL (dilution range 1:1-1:10), LDL/VLDL (dilution range 1:1-1:10) and free cholesterol (neat) was measure colourimetrically, in duplicates, after 40 minutes.
Signal from standard curve was measured colourimetrically over a period of time. Background signal subtracted and each point on the curve represents duplicate values (+/- SD).
Measurement of total cholesterol, HDL, LDL/VLDL from serum samples. Total Cholesterol (blue), HDL (green), and LDL/VLDL (cream) cholesterol were measured following the kit protocol.
This product has been referenced in:
- Suzuki K et al. Long-term oral administration of Exendin-4 to control type 2 diabetes in a rat model. J Control Release 294:259-267 (2019). Read more (PubMed: 30572033) »
- Yu J et al. Ezetimibe, a NPC1L1 inhibitor, attenuates neuronal apoptosis through AMPK dependent autophagy activation after MCAO in rats. Exp Neurol 307:12-23 (2018). Rat . Read more (PubMed: 29852178) »