Coenzyme II NADP+ or NADPH content testing kit and its method

A kit and reagent technology, applied in the field of life sciences, can solve problems such as non-universal use, high detection limit, and large interference of impurities

Inactive Publication Date: 2018-03-09
SUZHOU COMIN BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Coenzyme Ⅱ NADP + Or the determination method of NADPH content is glucose-6-phosphate dehydrogenase coupling method, and the shortcoming of this method is: 1, can not be universally used in animal, plant, microorganism and cultured cell sample; 2, detection limit is too high, can only reach 10 -3 mol / L; 3. Large interference by impurities, there are false positives; 4. The detection stability is not high

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  • Coenzyme II NADP+ or NADPH content testing kit and its method
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  • Coenzyme II NADP+ or NADPH content testing kit and its method

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Embodiment Construction

[0083] The present invention will be described in detail below in conjunction with embodiments.

[0084] A coenzyme II NADP + Or NADPH content determination kit, including the following reagents:

[0085] Acid extract, 50mL×1 bottle, made by distilling concentrated hydrochloric acid to 50mL, store at 4°C;

[0086] Alkaline extract, 50mL×1 bottle, made with NaOH to a volume of 50mL, stored at 4°C;

[0087] Reagent 1, liquid 15 mL×1 bottle, made by mixing HEPES and EDTA and distilling to 15 mL, adjusting the pH to 8.0 with KOH, and storing at 4°C;

[0088] Reagent 2, powder × 1 stick, composed of glucose 6-phosphate, placed in a 5mL reagent bottle, stored at -20°C;

[0089] Reagent 3, powder x 1, composed of PMS, placed in a 5mL brown bottle, stored at -20°C;

[0090] Reagent 4, powder × 1 tube, composed of MTT, placed in a 5mL brown bottle, stored at 4°C;

[0091] Reagent 5, liquid 1.8mL×1 tube, composed of 14U / mL glucose-6-phosphate dehydrogenase solution, placed in 2mL E...

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Abstract

The invention discloses a coenzyme II NADP+ or NADPH content testing kit and its method. The kit comprises acid extractive fluid, alkali extractive fluid, mixed solution of HEPES and EDTA, glucose 6-phosphate, PMS, MTT, glucose 6-phosphate dehydrogenase solution, NaCl solution and mixed solution of ethanol and double distilled water. The method includes steps of extracting NADP+ and NADPH in a sample by acid and alkali extracting fluids on the basis of a spectrophotometric method; performing PMS hydrogen delivering action on NADPH, and reducing MTT to be formazan; detecting light absorption value under 570 nm, and testing the NADPH content; reducing the NADP+ to be NADPH by glucose 6-phosphate dehydrogenase, thereby detecting the NADP+ content. The kit and its testing method can effectively test the content of the coenzyme II NADP+ or NADPH, and are featured by high specifity, wide application range, high stability, and others.

Description

technical field [0001] The invention belongs to the field of life sciences, in particular to a coenzyme II NADP + Or NADPH content assay kit and method thereof. Background technique [0002] Coenzyme II NADP(H) widely exists in animals, plants, microorganisms and cultured cells, and the determination of NADP + and NADPH content can be calculated as NADP (NADPH + NADP + ) content and NADPH / NADP + The ratio, its change is closely related to the pentose phosphate pathway and biosynthesis and antioxidant response. NADPH / NADP + The ratio is not only one of the main markers of the redox state of cells, but also plays an important regulatory role in the PPP pathway, biosynthesis and antioxidant metabolism. [0003] Coenzyme Ⅱ NADP + Or the determination method of NADPH content is glucose-6-phosphate dehydrogenase coupling method, and the shortcoming of this method is: 1, can not be universally used in animal, plant, microorganism and cultured cell sample; 2, detection limit ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31
CPCG01N21/31
Inventor 姚金美赵林川
Owner SUZHOU COMIN BIOTECH
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