(8 references)-
- 抗原
- Lipase
- 应用范围
- Activity Assay (AcA)
- 样品类型
- Serum, Plasma (no EDTA), Urine, Saliva
- 特异性
- 40 U/L
- 产品特性
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Sensitive and accurate. Linear detection range 40 to 1600 U/L lipase activity in 96-well plate assay.
Convenient and high throughput. The procedure involves adding a single working reagent, and reading the optical density at 10 min and 20 min at room temperature or 37°C. Can be automated to process thousands of samples per day. - 组件
- Assay Buffer (pH 8.5): 15 mL. Color Reagent: 530 mg. BALB Reagent: 1.0 mL. Calibrator: 2.0 mL (equivalent to 735 U/L).
- 试剂未包括
- Pipeting (multi-channel) devices. Clear-bottom 96-well plates (e.g. Corning Costar) and plate reader.
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- 应用备注
- Direct assays of lipase activity in serum, plasma, saliva, urine and other biological samples.
- 实验流程
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1. Transfer 150 µL H2O and 150 µL Calibrator into wells of a clear- bottom 96-well plate. Pipette 10 µL samples into separate wells. Add 140 µL Working Reagent to each sample well. Tap plate briefly to mix reaction mixture. Note: if the assay is to be performed at another temperature (e.g. 37°C), warm up the Working Reagent to this temperature prior to adding to the sample.
2. Read OD412nm on a plate reader at 10 min (OD10min) and at 20 min (OD20min). - 试剂准备
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Mix Color Reagent into Assay Buffer and shake vial to mix. Add 0.8 mL BALB Reagent (sufficient for 100 assays). Alternatively for partial reconstitution: for each well of reaction, mix 5 mg Color Reagent, 140 µL Assay Buffer and 8 µL BALB Reagent. The Working Reagent should be prepared freshly and used within one hour. Important: this assay is based on a kinetic reaction, addition of the Working Reagent should be quick. Use of a multi-channel pipettor is recommended.
- 样品制备
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Lipase inhibitors (EDTA, and certain detergents Tween-20, NP-40), mercaptoethanol and dithiothreitol interfere with this assay and should be avoided in sample preparation. Samples can be stored frozen for at least one month, if not assayed immediately. Tissue and cell lysates can be obtained by homogenization in cold PBS buffer and centrifugation (e.g. 5 min at 14,000 rpm).
- 限制
- 仅限研究用
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- 储存条件
- RT
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: "Macrophage-derived human resistin exacerbates adipose tissue inflammation and insulin resistance in mice." in: The Journal of clinical investigation, Vol. 119, Issue 3, pp. 531-9, (2011) (PubMed).
: "Effect of bile on the oral absorption of halofantrine in polyethylene glycol 400 and polysorbate 80 formulations dosed to bile duct cannulated rats." in: The Journal of pharmacy and pharmacology, Vol. 63, Issue 6, pp. 817-24, (2011) (PubMed).
: "Biochemical and histological effects of exendin-4 (exenatide) on the rat pancreas." in: Diabetologia, Vol. 53, Issue 1, pp. 153-9, (2010) (PubMed).
: "Arabidopsis N-MYC DOWNREGULATED-LIKE1, a positive regulator of auxin transport in a G protein-mediated pathway." in: The Plant cell, Vol. 21, Issue 11, pp. 3591-609, (2010) (PubMed).
: "Human risk allele HLA-DRB1*0405 predisposes class II transgenic Ab0 NOD mice to autoimmune pancreatitis." in: Gastroenterology, Vol. 139, Issue 1, pp. 281-91, (2010) (PubMed).
: "Caenorhabditis elegans dauers need LKB1/AMPK to ration lipid reserves and ensure long-term survival." in: Nature, Vol. 457, Issue 7226, pp. 210-4, (2009) (PubMed).
: "Methods for detection and characterization of lipases: A comprehensive review." in: Biotechnology advances, Vol. 27, Issue 6, pp. 782-98, (2009) (PubMed).
: "Pancreatic exocrine insufficiency in LXRbeta-/- mice is associated with a reduction in aquaporin-1 expression." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 105, Issue 39, pp. 15052-7, (2008) (PubMed).
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: "Macrophage-derived human resistin exacerbates adipose tissue inflammation and insulin resistance in mice." in: The Journal of clinical investigation, Vol. 119, Issue 3, pp. 531-9, (2011) (PubMed).
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- 抗原
- Lipase
- 别名
- EDL Kit, EL Kit, PRO719 Kit, 3110013K01Rik Kit, lipase Kit, mEDL Kit, lipase G, endothelial type Kit, lipase, endothelial Kit, LIPG Kit, Lipg Kit
- 背景
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Quantitative determination of lipase activity by colorimetric (412nm) method.
Procedure: 20 min.
Lipase catalyzes the hydrolysis of ester bonds on the glycerol backbone of a lipid substrate. In humans, pancreatic lipase is the key enzyme responsible for breaking down fats in the digestive system by converting triglycerides to monoglycerides and free fatty acids. Human pancreatic lipase and its related protein 2 are the main lipases secreted by the pancreas. In acute pancreatitis, lipase levels can rise 5 to 10-fold within 24 to 48 hours. Increased activities have also been associated with pancreatic duct obstruction, pancreatic cancer, kidney disease, salivary gland inflammation, bowel obstruction, and other pancreatic diseases. Decreased levels may indicate permanent damage to lipase-producing cells in the pancreas. Simple, direct and automation-ready procedures for measuring lipase activity are very desirable. This Lipase 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 colored product. The color intensity, measured at 412 nm, is proportionate to the enzyme activity in the sample.
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