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Kit for determining ATP content in trace amounts of sample and application thereof

A kit and a very small amount of technology, applied in the direction of measuring devices, instruments, fluorescence/phosphorescence, etc., can solve the problems of no suitable and feasible methods, achieve accurate results, simple operation, and avoid time-consuming effects

Inactive Publication Date: 2016-05-04
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although existing methods and commercial kits can detect ATP content in tissues and a large number of cells, there is no suitable and feasible method for precious and extremely small samples such as oocytes and early embryos.

Method used

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  • Kit for determining ATP content in trace amounts of sample and application thereof
  • Kit for determining ATP content in trace amounts of sample and application thereof
  • Kit for determining ATP content in trace amounts of sample and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Related experiments were carried out to determine the ATP content in mouse oocytes.

[0032] 1. Sample Preparation

[0033] ICR 3-week-old mouse oocytes were selected as samples for the experiment. After the mice were sacrificed by cervical dislocation, the GV stage oocytes were collected in a sterile 35mm culture dish for use.

[0034] 2. Determination of ATP content

[0035] 1) Use currently commercially available kits to detect the ATP content in different numbers of oocytes

[0036] The kit contains ATP detection mixed powder, containing luciferase, luciferin, magnesium sulfate, dithiothreitol, ethylenediaminetetraacetic acid, bovine serum albumin, tris (hydroxymethyl) methyl buffer salt, plus 50mL Ultrapure water until dissolved and ready for use; ATP analysis mixed powder, containing magnesium sulfate, dithiothreitol, ethylenediaminetetraacetic acid, bovine serum albumin, tris (hydroxymethyl) methyl buffer salt, add 5mL ultrapure water , vortex until dissolved,...

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PUM

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Abstract

The invention relates to a kit for determining ATP content in trace amounts of sample and application thereof. The kit comprises a solution I which is a mixture of TrisHCl and bovine serum albumin; a solution II which is a sodium hydroxide solution; a solution III which is a mixture of hydrochloric acid and the TrisHCl; a reagent I comprising the TrisHCl, the bovine serum albumin, magnesium chloride, dithiothreitol, D-glucose, nicotinamide adenine dinucleotide phosphate, hexokinase and colourless glucose-6-phosphate dehydrogenase; a reagent II comprising Imidapril HCl, the bovine serum albumin, alpha-ketoglutaric acid monosodium salt, sodium hydroxide, ammonium acetate, adenosine diphosphate, glucose-6-phosphoric acid, glutamate dehydrogenase and the colourless glucose-6-phosphate dehydrogenase; a reagent III comprising the Imidapril HCl, ethylenediaminetetraacetic acid, the magnesium chloride, the ammonium acetate, NADP and glucose 6-phosphate dehydrogenase. The kit is simple to operate, and the determination result is more accurate.

Description

technical field [0001] The invention belongs to the technical field of ATP content detection, and in particular relates to a kit for measuring ATP content in very small samples and an application thereof. Background technique [0002] In the process of energy conversion and transfer in organisms, 5'-adenosine 5'-triphosphate (ATP) is a key substance. Mitochondria is an important place for ATP synthesis, and the ATP produced by it is the main energy source for cell life activities. ATP is closely related to the functional state of mitochondria, and as a key energy molecule, it plays an important role in various physiological and pathological processes of cells. Changes in ATP levels can affect many cellular functions. Usually, the ATP content will decrease when the cells are in apoptosis, necrosis or in some toxic state; however, factors such as high glucose stimulation may up-regulate the ATP content in some types of cells. Therefore, accurate determination of ATP levels ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 王强葛娟侯晓静
Owner NANJING MEDICAL UNIV
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