Description
Cholesterol Assay Kit (BA0064) (BA0064)
The Cholesterol Assay Kit (SKU: BA0064) provides a simple, direct and automation-ready procedure for measuring total cholesterol. Cholesterol is a sterol and lipid present in cell membranes and transported in the bloodstream of all animals, playing important roles in forming membranes and hormones and in cell signalling. Elevated levels (hypercholesterolaemia) are associated with cardiovascular diseases such as atherosclerosis, whereas low levels (hypocholesterolaemia) may be linked to depression, cancer and cerebral haemorrhage. This assay uses a single working reagent that combines cholesterol ester hydrolysis, oxidation and colour reaction in one step. The colour intensity at 570 nm, or fluorescence intensity at excitation/emission 530/585 nm, is directly proportional to total cholesterol concentration. The room-temperature protocol requires no 37C heater and is readily automated for high-throughput work.
| Product Name: | Cholesterol Assay Kit (BA0064) |
| SKU: | BA0064 |
| Detection Method: | Colorimetric or Fluorometric |
| Detection Range: | Colorimetric 0.1 to 10 mg/dL; fluorometric 0.02 to 2 mg/dL |
| Sample Type: | Serum, plasma and other biological samples |
| Species Reactivity: | All |
| Assay Time: | 30 minutes |
| Kit Size: | 100 Assays |
| Equipment Required: | Microplate reader |
| Storage: | -20C |
| Shelf Life: | 12 months after receipt |
| Shipping: | Gel Pack |
Quantitative colorimetric or fluorometric determination of total cholesterol. A single working reagent combines cholesterol ester hydrolysis, oxidation and colour reaction in one step, read at 570 nm or fluorometrically at excitation/emission 530/585 nm.
- Sensitive and accurate
- Colorimetric linear detection range 0.1 to 10 mg/dL; fluorometric 0.02 to 2 mg/dL
- Convenient room-temperature assay with no 37C heater required
- High-throughput and readily automated
- Direct assays of cholesterol in serum, plasma and other biological samples
- Pharmacology: effects of drugs on cholesterol metabolism
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
| Step | Procedure |
| 1 | Standard curve. Bring all reagents to room temperature. Prepare a 10 mg/dL standard by mixing 15 uL 300 mg/dL Standard with 435 uL Assay Buffer, then prepare the dilution series shown in the table. Transfer 50 uL diluted standards into a clear 96-well plate and 50 uL diluted sample into separate wells. |
| 2 | For each reaction well, mix 55 uL Assay Buffer with 1 uL Enzyme Mix and 1 uL Dye Reagent. Add 50 uL of this working reagent to each standard and sample well and tap to mix. |
| 3 | Incubate 30 minutes at room temperature and read OD at 570 nm. If the sample OD exceeds the 10 mg/dL standard, dilute in assay buffer and repeat. |
| 4 | Fluorimetric procedure. Dilute the standards 1:10 in Assay Buffer, transfer 50 uL standards and 50 uL samples into a black 96-well plate, add 50 uL working reagent, tap to mix, incubate 30 minutes at room temperature and read fluorescence at excitation 530 nm and emission 585 nm. |
Subtract the blank OD (water, standard #8) from the standard OD values and plot against standard concentrations to determine the slope by linear regression. The cholesterol concentration of the sample is calculated from the standard curve and multiplied by the dilution factor n (generally 10-30 for blood samples).
| Component | Quantity | Storage |
| Assay Buffer | 20 mL | -20C |
| Dye Reagent | 120 uL | -20C |
| Enzyme Mix | 120 uL | -20C |
| Standard (300 mg/dL cholesterol) | 1 mL | -20C |
