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Cell proliferation and toxicity detection kit

A technology for toxicity detection and cell proliferation, which is applied in the field of cell biology, can solve the problems of no reaction termination reagent, uncontrollable reaction time, leakage or overfilling of detection solution, etc., to save time, stabilize the preservation environment, and avoid deterioration.

Active Publication Date: 2019-03-01
大连博格林生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the kit based on WST-8 detection still has the following defects: the detection solution containing PMS is the same pink color as the culture medium, which may easily cause leakage or overfilling of the detection solution; there is no matching reaction termination reagent, and the reaction time cannot be controlled. It is not suitable for large-scale drug screening; the stability of the detection solution is poor, and it is easy to deteriorate during storage or transportation at high temperature; the reaction time and detection sensitivity need to be further improved

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] 1. The preparation method of 100 usages of the Cell Proliferation and Toxicity Detection Kit:

[0018] Preparation of detection solution: Weigh 8.6 mg of sodium chloride, 0.3 mg of potassium chloride, and 0.28 mg of calcium chloride and fully dissolve them in 1 ml of ultrapure water, then add 7.8 mg of WST-8 and 0.138 mg of 2-methyl -1,4-Naphthoquinone, fully dissolved and sterilized through a 0.22μm filter membrane, packed in a sterilized 1.5 ml Keli brown tube.

[0019] 2. Preparation of reaction termination solution: Add 8.5 μl HCl to 991.5 μl ultrapure water, mix well, pass through a 0.22 μm filter membrane to sterilize, and put it in a 1.5 ml standable transparent tube.

[0020] 3. The above two reagents pass the sterility test and are packed in the kit at a ratio of 1:1, which constitutes the cell proliferation and toxicity detection kit.

[0021] The method for counting C6 cells, detecting cell activity and proliferation using the kit of the embodiment of the pr...

Embodiment 2

[0028] The preparation method of 100 usages of the cell proliferation and toxicity detection kit:

[0029] 1. Preparation of test solution: Weigh 8.6 mg of sodium chloride, 0.3 mg of potassium chloride, and 0.28 mg of calcium chloride and dissolve them in 1 ml of ultrapure water, then add 7.2 mg of WST-8 and 0.12 mg of 2- Methyl-1,4-naphthoquinone, fully dissolved, sterilized through a 0.22 μm filter membrane, and dispensed into sterilized 1.5 ml standable brown tubes.

[0030] 2. Preparation of reaction termination solution: Add 9 μl HCl to 991 μl ultrapure water, mix well, pass through a 0.22 μm filter membrane to sterilize, and dispense into 1.5 ml clear tubes.

[0031] 3. The above two reagents have passed the sterility test and are packed in the kit at a ratio of 1:1 to form a cell proliferation and toxicity detection kit.

[0032] Use the embodiment 2 kit of the present invention to carry out the toxicity detection method of paclitaxel to Hela cell:

[0033] 1. Prepare...

Embodiment 3

[0045] The preparation method of 500 usages of the Cell Proliferation and Toxicity Detection Kit:

[0046] 1. Preparation of test solution: Weigh 43 mg of sodium chloride, 1.5 mg of potassium chloride, and 1.4 mg of calcium chloride and dissolve them in 5 ml of ultrapure water, then add 33 mg of WST-8 and 0.52 mg of 2- Methyl-1,4-naphthoquinone, fully dissolved, sterilized through a 0.22μm filter membrane, and dispensed into sterilized 8 ml brown liquid bottles.

[0047] 2. Preparation of reaction termination solution: Add 42.5 μl HCl into 4.96 ml ultrapure water, mix well, pass through a 0.22 μm filter membrane to sterilize, and dispense into 8 ml natural color liquid bottles.

[0048] 3. The above two reagents pass the sterility test and are packed in the kit at a ratio of 1:1, which constitutes the cell proliferation and toxicity detection kit.

[0049] The method for detecting suspension cell activity and proliferation using the kit of Example 3 of the present invention: ...

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Abstract

The invention discloses a cell proliferation and toxicity detection kit. The cell proliferation and toxicity detection kit has the advantages that the sensitivity and stability of detection can be improved, the reaction time is shortened, and the omitted adding of a detection solution is avoided. The cell proliferation and toxicity detection kit comprises the detection solution and a reaction terminating solution, wherein the detection solution is prepared from WST-8, 2-methyl-1,4-naphthoquinone, an aqueous solution of sodium chloride, an aqueous solution of potassium chloride and an aqueous solution of calcium chloride, the mole ratio of the WST-8 to the 2-methyl-1,4-naphthoquinone is (12 to 18): 1, concentrations of aqueous solutions of sodium chloride, potassium chloride and calcium chloride separately are 8.6g / L, 0.3g / L and 0.28g / L; the reaction terminating solution is HCl of 0.05mol / L to 0.15mol / L.

Description

technical field [0001] The invention relates to the technical field of cell biology, in particular to a cell proliferation and toxicity detection kit which can improve detection sensitivity and stability, shorten reaction time, and avoid missed addition of detection solution. Background technique [0002] Cell proliferation and toxicity detection is a basic method for determining the toxicity of substances, evaluating drug safety and cell health, and can also indicate the proliferation activity of tumor cells, the cell proliferation activity of target gene transient / stable cell lines, the overexpression of target genes or RNAi interference. Gene function research plays an important role in drug screening and functional safety research. Therefore, cell proliferation and toxicity detection technology has been widely used in various research fields such as molecular biology, tumor biology, immunology, genetics, pharmacology and pharmacokinetics. [0003] At present, cell proli...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/52
CPCG01N33/52
Inventor 姜梦李民强
Owner 大连博格林生物科技有限公司
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