FLAG fusion tag colloidal gold test strip and preparation method thereof

A technology that integrates tags and colloidal gold, used in biological research and biological fields, can solve the problems of high price and high requirements, and achieve the effect of low cost, simplified operation, and small amount of reagents and samples.

Inactive Publication Date: 2010-05-12
河南省生物工程技术研究中心
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Problems solved by technology

The former requires expensive fully automatic instruments and various kits that are strictly matched with the instruments, and is subject to higher requirements

Method used

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Examples

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Embodiment Construction

[0014] 1. Preparation of 12-30nm colloidal gold by trisodium citrate method Take a 250ml Erlenmeyer flask, add 100ml double distilled water and 1ml 1% gold chloride, heat to boil; add 1.5ml 1% sodium citrate to the above solution. Mix well, and keep boiling for 30 minutes, the color of the solution first turns black, then gradually turns red.

[0015] 2. Determination of the optimum pH for the combination of protein and colloidal gold

[0016] (1) Take several 1.5ml test tubes and add 1ml 10nm colloidal gold respectively;

[0017] (2) Use 25mM K2CO3 to adjust the pH to 3, 4, 5, 6, 7, 8, 9, 10;

[0018] (3) Take a 96-well culture plate, add 100ul of the above-mentioned colloidal gold into the wells according to the pH from low to high, and repeat three times;

[0019] Add 3ul of Flag antibody with a concentration of 1mg / ml to each well, mix, and place at room temperature for 10-15min;

[0020] Add 20ul of 10% NaCl solution to each well, mix, and place at room temperature for...

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Abstract

The invention relates to a preparation method of a colloidal gold reagent and a rapid detection method for a fusion tag. The colloidal gold reagent belongs to a detection reagent of a FLAG fusion tag in the field of biologic research. A double- antibody sandwich method immunoassay principle is adopted in the method, and a Flag monoclonal antibody and a Flag polyclonal antibody which are purified are respectively used as a colloidal gold combined antibody and an envelope antibody, thus the gene engineering expression protein adopting FLAG as the fusion tag is rapidly detected.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular, relates to the preparation of a colloidal gold reagent and the rapid detection of fusion tags, and belongs to the detection reagent of FLAG fusion tags in the field of biological scientific research. Background of the invention [0002] The fusion tag is a short peptide (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) containing eight amino acids, which is widely used as a fusion tag in the expression of genetically engineered proteins. With the increasing application of immunoassays, immunoassays are developing in two directions: one is fully automated immunoassays, and the other is rapid immunoassays using nitrocellulose membranes as carriers. The former requires expensive fully automatic instruments and various kits that are strictly matched with the instruments, and is subject to higher requirements. With the needs of convenient, rapid and popular detection, the immunocolloidal gold rapid diagno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/532G01N33/68G01N33/558
Inventor 王云龙李玉林李恒思王继美王国强
Owner 河南省生物工程技术研究中心
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