Kit for detecting lipase by enzyme method and preparation method
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A kit and lipase technology, applied in the field of enzymatic detection lipase kit and its preparation, to achieve the effects of good detection effect, strong protective effect, and obvious protective effect
Inactive Publication Date: 2013-06-26
SHANGHAI FOSUN PHARMA (GROUP) CO LTD +1
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Abstract
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[0003] The technical problem to be solved by the present invention is to overcome above-mentioned deficiencies, research and de
Method used
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Embodiment 1
[0024] Embodiment 1 prepares lipase detection kit
[0025] composition:
[0026] Reagent 1: (2L) (reagent volume)
[0027]
[0028] Deionized water is added to a total volume of 2L;
[0029] Reagent 2: (1L) (reagent volume)
[0030]
[0031]
[0032] Deionized water was added to a total volume of 1 L.
[0033] preparation:
[0034] (1) Preparation of reagent 1: (2L)
[0035] (1) First add 1.6L of deionized water to the container;
[0036] (2) Add TRIS buffer solution, sodium deoxycholate, calcium chloride, and sodium azide in sequence;
[0037] (3) Add enzyme: colipase
[0038] (4) Finally, add deionized water until the total volume is 2L and mix evenly to obtain;
[0039] (2) Preparation of reagent 2: (1L)
[0040] Add tartaric acid, taurodeoxycholic acid, mannitol proclin300 (biological preservative), thioglycolic acid, 6-methyl resorufin, and deionized water to the container in sequence until the total volume is 2 L, and mix well.
Embodiment 2
[0042] Reagent 1: (2L)
[0043]
[0044] Deionized water was added to a total volume of 2 L.
[0045] Reagent 2: (1L)
[0046]
[0047] Deionized water was added to a total volume of 1 L.
Embodiment 3
[0049] Reagent 1: (2L)
[0050]
[0051] Deionized water was added to a total volume of 2 L.
[0052] Reagent 2: (1L)
[0053]
[0054] Deionized water was added to a total volume of 1 L.
[0055] Preparation method is the same as embodiment 1
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Abstract
The invention provides a kit for detecting lipase by an enzyme method, and is composed of a reagent 1 and a reagent 2. The reagent 1 is composed of 10-200mmol/L of 2L buffer, 0.01-20ml/L of sodium deoxycholate, 0.1-2mg/L of colipase, 5-20mmol/L of calcium chloride, 0.5-2g/L of sodium azide, and the balance of deionized water to 2L; and the reagent 2 comprises the following components by weight: 10-200mmol/L of 1L tartrate, 0.1-10mmol/L of bezoar sodium deoxycholate, 10-200mmol/L of mannitol, 0.01-0.5ml/L of proclin300, 10-200mmol/L of mercaptoacetic acid, 0.1-0.5mmol/L of 6-methyl resorufin, and the balance of deionized water to 1L. The kit has strong protection effect on a substrate, and the stability of the reagent is increased, the kit has good detection effect, and has large clinical application value.
Description
technical field [0001] The invention relates to biological reagents, in particular to a detection kit, in particular to an enzymatic lipase detection kit and a preparation method thereof. Background technique [0002] Lipase (Lipase EC3.1.1.3., LPS, glyceride hydrolase) is a class of enzymes that hydrolyze oil and ester. The hydrolysis substrate of lipase is generally natural oil, and its hydrolysis site is the ester bond connecting fatty acid and glycerol in oil. It is different from other hydrolases. The lipase catalytic system is a heterogeneous system. The water-soluble enzyme catalysis occurs at the interface between the water-insoluble substrate and water. The catalytic mechanism of this interface is not very clear. The reaction mechanism between enzyme and substrate cannot be simply explained by Michaelis-Menten theory. Some people put forward the hypothesis that lipase is localized on the oil-water interface, and put forward a model of "ultrasubstrate", which can e...
Claims
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