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A kind of serum diluent in elisa test

A diluent and serum technology, applied in the field of serum diluent, can solve the problems of deep background color, high OD value, affecting the judgment of ELISA test results, etc., and achieve the effect of simple process operation and low equipment requirements

Active Publication Date: 2016-08-24
YEBIO BIOENG OF QINGDAO
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the ELISA test where the protein expressed by the vector is used as the coating antigen, although the antigen is purified and the purity is high, it often occurs that the background color is darker and the OD value is higher, which directly affects the judgment of the ELISA test result.

Method used

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  • A kind of serum diluent in elisa test
  • A kind of serum diluent in elisa test

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] 1. Preparation of diluent

[0013] 1.1 Preparation of Escherichia coli BL21 (E.coli BL21) bacterial liquid:

[0014] Pick a single colony of Escherichia coli BL21 (E.coli BL21) from the LB culture plate, transfer it to a test tube containing LB culture solution, and culture it overnight at 37°C to prepare a saturated bacterial solution. Transfer the saturated bacterial solution to 250ml of LB medium with an inoculum size of about 5%, and cultivate for 8 hours. Centrifuge at 6000 rpm for 15 minutes. The cells were dissolved in 15ml PBS and stored at -20°C.

[0015] 1.2 Extraction of soluble protein from Escherichia coli BL21 (E.coli BL21) Thaw the frozen bacterial solution and lyse it by ultrasonic. Set at 6000 rpm, centrifuge for 15 minutes, and collect the supernatant. Protein content was determined with BCA protein content detection kit. Then the bacteriostasis solution was diluted with 0.01M pH7.2PBS to a total protein content of 1.2mg / ml.

[0016] The specific...

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Abstract

The invention provides a serum diluent in ELISA detection. A primary antibody dilution liquid in an ELISA antibody detection test is mainly used for diluting a primary antibody in the ELISA test, and can prevent non-specific reaction in the ELISA test, reduce an OD value of low negative background and improve the sensitivity of the ELISA test. The serum diluent is prepared from fermented liquid of colon bacillus. According to the diluent, the problems of high negative background and low sensitivity in the ELISA test process are solved. The process is easy to operate, the requirement on the equipment is low, and massive production of the serum diluent can be achieved.

Description

technical field [0001] The invention belongs to the technical field of molecular detection, and in particular relates to a serum diluent in ELISA detection. Background technique [0002] In conventional ELISA tests, PBST, PBS containing 1% BSA, etc. are mostly used as serum diluents. In the ELISA test where the protein expressed by the vector is used as the coating antigen, although the antigen is purified and the purity is high, it often occurs that the background color is darker and the OD value is higher, which directly affects the judgment of the ELISA test result. . Therefore, it is necessary to provide a serum diluent with better effect. Contents of the invention [0003] The purpose of the present invention is to provide a serum diluent in the ELISA detection, i.e. the primary antibody (pig serum or chicken serum) dilution in the ELISA antibody detection test, which is mainly used in the primary antibody (pig serum or chicken serum) in the ELISA test. Serum) dilu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/531G01N33/96
CPCG01N33/531G01N33/54393
Inventor 徐保娟荣俊刘蕾郭伟伟宫晓朱艳梅王丽萍李陆梅孙健胡潇王树明宋新宇于晓云
Owner YEBIO BIOENG OF QINGDAO