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Dot blot hybridization reaction device and application thereof, and immunoblotting detection method

A reaction device and dot hybridization technology, applied in biochemical equipment and methods, measuring devices, biochemical cleaning devices, etc., can solve the problems of cross-contamination between samples, time-consuming and laborious operation, and difficulty in standardization, etc. Contamination, the effect of easy and rapid detection

Pending Publication Date: 2019-03-08
天康生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The existing dot hybridization technology is time-consuming and laborious to operate, difficult to standardize, easy to cause cross-contamination between samples, and is not suitable for operation in clinical sites other than laboratories. Therefore, an improved dot hybridization reaction device is currently in demand in the market

Method used

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  • Dot blot hybridization reaction device and application thereof, and immunoblotting detection method
  • Dot blot hybridization reaction device and application thereof, and immunoblotting detection method
  • Dot blot hybridization reaction device and application thereof, and immunoblotting detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] This embodiment provides a dot blot reaction device, refer to figure 1 , figure 2 and image 3 shown. The dot blot reaction device provided in this embodiment includes a sample loading and diafiltration module 100. The sample loading and diafiltration module 100 includes: a microwell sample loading plate 110, on which a sample through hole 111 and a frame flange 112 are arranged on the microwell sample loading plate. The diafiltration module 100 also includes a hybridization membrane layer 120, a filter paper layer 130, a gasket layer 140 and a support layer 150, and an annular flange 151 is arranged on the support layer 150; The filter tank body 160 is provided with a groove 161, and the side wall of the suction filter tank body 160 is provided with a suction filter hole 162;

[0084] The dot blot reaction device provided in this embodiment also includes a suction filtration module: comprising an oil-free vacuum pump 210, a pressure gauge 220 and a suction filtrati...

Embodiment 2

[0095] In this example, the dot blot reaction device provided in Example 1 was used to measure the content of E2 recombinant protein in the production process of CSFV E2 protein recombinant baculovirus inactivated vaccine. Proceed as follows:

[0096] (i) Preparation and pretreatment of the hybridization membrane layer 120: take a cut 10.5cm×7.5cm PVDF membrane as the hybridization membrane layer 120, activate it with methanol for 10s, soak it in the wet transfer buffer, and prepare a thin filter paper pad of the same size Below the gasket layer 140 to prevent leakage.

[0097] (ii) Assembling the sample-loading diafiltration module 100: install from bottom to top according to the order of suction filtration tank body 160-support layer 150-sealing gasket layer 140-filter paper layer 130-hybridization membrane layer 120-micropore sample loading plate 110, After each component is fixed, it is placed in a constant temperature incubation tank 300 and connected to an oil-free vacu...

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Abstract

The invention provides a dot blot hybridization reaction device and application thereof, and an immunoblotting detection method and relates to the technical field of molecular hybridization experimental devices. The dot blot hybridization reaction device comprises a sampling infiltration module and a suction filtration module used for supplying a negative pressure environment for the interior of the sampling infiltration module, wherein the sampling infiltration module comprises a micropore sampling plate provided with a sample through hole, a support layer provided with an infiltration through hole corresponding to the sample through hole, and a suction filtration tank; a first actuation unit is arranged in the micropore sampling plate; a second actuation unit matched with the first actuation unit is arranged in the suction filtration tank; a closed inner chamber is formed by the matched actuation units in the manner of adsorbing and fixing; and a hybridization film layer and the support layer are successively fixed under the micropore sampling plate. The dot blot hybridization reaction device is portable, is simple in operation and can effectively prevent the problem of cross contamination of sample rooms of the dot blot hybridization reaction device.

Description

technical field [0001] The invention relates to the technical field of molecular hybridization experimental devices, in particular to a dot hybridization reaction device and its application and immunoblotting detection method. Background technique [0002] Molecular hybridization technology is a commonly used detection method for biological macromolecules such as nucleic acids and proteins. Nucleic acid molecular hybridization is a method of using labeled DNA or RNA as a probe to detect a specific nucleic acid sequence; protein molecular hybridization is the application of specific binding of antigens and antibodies. The principle is to detect specific polypeptides and proteins by using a chromogenic agent labeled with an antibody through a chemical reaction, and molecular hybridization has high sensitivity and high specificity. [0003] Dot hybridization is a method in molecular hybridization technology, which usually includes the following steps: firstly, the hybridization...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12M1/38C12Q1/6834G01N33/53
CPCG01N33/5304C12Q1/6834C12Q2565/631
Inventor 贺笋王遵宝徐龙飞潘晓梅李俊辉李延涛候玉珍田晓艳
Owner 天康生物制药有限公司
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