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| Product Name | Lipase Enzyme Activity Detection Kit (Colorimetric) |
| Catalog No. | ETR-HMM-0067 |
| Description | A colorimetric assay kit for quantifying lipase enzyme activity using 1,2-diacylglycerol or para-nitrophenyl palmitate (pNP-palmitate) as substrates. Lipase hydrolyzes the ester bonds in these substrates, releasing the chromogenic para-nitrophenol group, which is measured at 405 nm in alkaline conditions. |
| Intended Use | Measurement of lipase activity in serum and plasma for pancreatitis biomarker research, evaluation of microbial lipase production in fermentation biotechnology, and quality control of lipase enzyme preparations for industrial applications. |
| Principle / Technology | Lipase (EC 3.1.1.3) cleaves the sn-1 or sn-3 ester bonds of glycerophospholipids and triglycerides; the synthetic substrate para-nitrophenyl palmitate (pNP-palmitate) is an emulsified ester that is hydrolyzed at the fatty acid ester bond, releasing palmitate and para-nitrophenol (pNP); pNP is yellow at alkaline pH with molar absorptivity 18,000 M-1cm-1 at 405 nm; activity is proportional to pNP release rate. |
| Detection Method | Colorimetric absorbance at 405 nm using a microplate spectrophotometer; substrate must be used as an emulsion to provide an aqueous-lipid interface |
| Sample Type | Serum, plasma, cell culture media from pancreatic cells, pancreatic tissue homogenates, food samples, bacterial and fungal enzyme preparations, industrial lipase solutions |
| Performance Range / Specifications | Activity range: 0.5–500 U/L for serum lipase; one unit = amount releasing 1 μmol pNP per minute at 37°C; substrate emulsion must be freshly prepared before each assay; linear reaction rate over 5–10 minutes |
| Sensitivity / LOD | Detection limit: approximately 0.2 U/L for serum lipase with 5 μL sample; requires substrate emulsification for triglyceride-mimicking interface |
| Specificity | Pancreatic lipase specifically hydrolyzes at the sn-1 and sn-3 positions; phospholipase activity and non-specific esterases contribute minimally under the substrate and buffer conditions; colipase cofactor is not required for pNP-palmitate hydrolysis but is required for triglyceride substrates in physiological assays |
| Reaction Conditions / Protocol | Prepare pNP-palmitate emulsion (2 mM pNP-palmitate in Tris buffer pH 8.0 with 0.5% gum arabic, sonicate or vortex vigorously until stable); warm Emulsion to 37°C; add 5–10 μL diluted sample to 200 μL substrate emulsion in microplate; mix by brief pipetting; monitor absorbance at 405 nm kinetically at 37°C every 1 minute for 10 minutes; calculate initial linear rate; compare to pNP standard curve for activity calculation |
| Components / Formulation | pNP-Palmitate Substrate (lyophilized, 100 mg), Gum Arabic Emulsifier (10% w/v solution), Tris-HCl Assay Buffer pH 8.0, CaCl2 cofactor (100 mM), pNP Standard Solution (for activity unit calibration), Lipase Positive Control (porcine pancreatic lipase, lyophilized) |
| Storage Conditions | pNP-palmitate lyophilized: -20°C desiccated for 24 months; pNP Standard: -20°C for 12 months; buffers at 2–8°C; porcine lipase control at -20°C |
| Shelf Life | 24 months for lyophilized substrate |
| Package Specifications | 100 assays, 500 assays (96-well microplate format) |
| Product Form | Lyophilized substrate and control with liquid buffer and emulsifier |
| Quality Control | Each lot tested with porcine pancreatic lipase reference; specific activity within ±20% of nominal; emulsion preparation reproducibility confirmed; pNP standard curve R2 ≥0.999 |
| Key Features | pNP-palmitate is the most widely used lipase substrate for activity assays; gum arabic emulsifier creates stable long-chain substrate interface; compatible with serum lipase measurement relevant to pancreatitis clinical research; positive control included for inter-run normalization |
For research use only, not for clinical use.
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