Cell Biology Assays
Lipase Activity Assay Kit (Colorimetric) (BA0052)
- SKU:
- BA0052
- Product Type:
- Assay
- Detection Method:
- Colorimetric
- Instrument:
- Microplate Reader
- Sample Type:
- Serum, Plasma, Saliva, Urine
- Research Area:
- Lipid Metabolism
- Research Area:
- Diabetes & Obesity
- Research Area:
- Plant & Environmental Stress
- Research Area:
- Enzyme Activity
- Research Area:
- Clinical Chemistry
Description
Lipase Activity Assay Kit (Colorimetric) (BA0052) (BA0052)
The Lipase Activity Assay Kit (Colorimetric) (SKU: BA0052) provides a simple, direct and automation-ready method for measuring lipase activity in serum, plasma, saliva, urine and other biological samples. Lipase catalyses the hydrolysis of ester bonds on the glycerol backbone of lipid substrates, and pancreatic lipase is the key enzyme responsible for breaking down fats in the digestive system. In acute pancreatitis, lipase levels can rise 5- to 10-fold within 24 to 48 hours, and raised activity is also associated with pancreatic duct obstruction, pancreatic cancer, kidney disease and bowel obstruction. The assay is based on an improved dimercaptopropanol tributyrate (BALB) method, in which SH groups formed from lipase cleavage of BALB react with 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) to form a yellow-coloured product. The colour intensity, measured at 412nm, is proportional to the enzyme activity in the sample.
| Product Name: | Lipase Activity Assay Kit (Colorimetric) (BA0052) |
| SKU: | BA0052 |
| Detection Method: | Colorimetric (kinetic) |
| Detection Range: | 40 to 1600 U/L lipase activity in 96-well plate assay |
| Sample Type: | Serum, plasma, saliva, urine and other biological samples |
| Species Reactivity: | All |
| Assay Time: | 20 minutes (readings at 10 and 20 minutes) |
| Kit Size: | 100 Assays |
| Equipment Required: | Microplate reader |
| Storage: | The kit is shipped at room temperature. Store all components at 4°C. |
| Shelf Life: | 12 months after receipt |
| Shipping: | Room Temperature |
This kit is based on an improved dimercaptopropanol tributyrate (BALB) method, in which SH groups formed from lipase cleavage of BALB react with DTNB to form a yellow-coloured product. The colour intensity, measured at 412nm, is proportional to the enzyme activity in the sample.
- Sensitive and accurate: linear detection range 40 to 1600 U/L lipase activity in 96-well plate assay
- Convenient and high-throughput: addition of a single working reagent and reading the optical density at 10 and 20 minutes at room temperature or 37°C, automatable for thousands of samples per day
- Direct assays of lipase activity in serum, plasma, saliva, urine and other biological samples
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 | Prepare Working Reagent by mixing the Color Reagent into the Assay Buffer and shaking, then adding 0.8 mL BALB Reagent (sufficient for 100 assays). Prepare fresh and use within one hour. For partial reconstitution, per well mix 5 mg Color Reagent, 140 µL Assay Buffer and 8 µL BALB Reagent. |
| 2 | Transfer 150 µL H2O and 150 µL Calibrator into wells of a clear-bottom 96-well plate. Pipette 10 µL samples into separate wells and add 140 µL Working Reagent to each sample well; tap plate briefly to mix. Warm the Working Reagent to the assay temperature (e.g. 37°C) beforehand if required. |
| 3 | Read OD412nm on a plate reader at 10 minutes (OD10min) and at 20 minutes (OD20min). |
Lipase activity = [(OD20min − OD10min) / (ODCalibrator − ODH2O)] × 735 (U/L), where OD20min and OD10min are the OD412nm values of the sample at 20 and 10 minutes, and ODCalibrator and ODH2O are the OD412nm values of the Calibrator and water at 20 minutes. If the calculated activity exceeds 1600 U/L, dilute the sample in water, repeat the assay and multiply by the dilution factor. Unit definition: one unit of enzyme catalyses the cleavage of 1 µmole of substrate per minute at pH 8.5.
| Component | Quantity | Storage |
| Assay Buffer (pH 8.5) | 15 mL | 4°C |
| Color Reagent (BALB) | 530 mg | 4°C |
| BALB Reagent | 1.0 mL | 4°C |
| Calibrator (equivalent to 735 U/L) | 2.0 mL | 4°C |
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