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Method for determining ribosome inactivating protein activity

A technology of ribosome inactivation and protein activity, applied in the field of biochemical analysis and detection, can solve problems such as poor quantitative effect and inconvenience, and achieve the effect of wide practical application prospect, sensitive response, and avoidance of interference.

Active Publication Date: 2017-02-22
CHENGDU MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method can be accurately quantified, but it also needs to exclude the interference of RNase, and RP-HPLC can only measure one sample at a time, which is inconvenient if there are a large number of samples to be measured
[0004] Patent Publication No. 104498494A discloses a method for detecting RIP activity using plasmids, but this method needs to detect RIP activity by agarose gel electrophoresis, and the quantitative effect is not good

Method used

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  • Method for determining ribosome inactivating protein activity
  • Method for determining ribosome inactivating protein activity
  • Method for determining ribosome inactivating protein activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Embodiment 1 The preparation of colored liquid of the present invention

[0081] The chromogenic solution used in the present invention is a coupling enzyme reagent for detecting the content of adenine. The chromogenic solution is prepared with fresh ultrapure water before use. It is a colorless and transparent solution and stored at 4°C. If it contains a stabilizer, the validity period is at least 3 months. . If it is found that the light absorption value of the blank tube is greater than 0.01 during the measurement, it should be re-prepared.

[0082] The coupled enzyme chromogenic solution was prepared as follows:

[0083] Take 0.1M, pH7.0 MES-NaOH buffer, add TOOS, 4-aminoantipyrine, peroxidase, adenosine deaminase, xanthine oxidase in turn, and finally add MES-NaOH buffer to set Make the chromogenic solution contain TOOS at a final concentration of 40mM, 4-aminoantipyrine at 5g / L, peroxidase at 9U / mL, adenosine deaminase at 6U / mL, yellow Purine oxidase. It is th...

Embodiment 2

[0084] Embodiment 2 Preparation of colored liquid of the present invention

[0085] Take 0.1M, pH7.0 MES-NaOH buffer, add TOOS, 4-aminoantipyrine, peroxidase, adenosine deaminase, xanthine oxidase in turn, and finally add MES-NaOH buffer to set Make the chromogenic solution contain TOOS at a final concentration of 35mM, 4-aminoantipyrine at 4g / L, peroxidase at 10U / mL, adenosine deaminase at 5U / mL, yellow Purine oxidase. It is the chromogenic solution of the present invention, which is placed in a brown bottle and stored at 4°C for future use.

Embodiment 3

[0086] Embodiment 3 The preparation of colored liquid of the present invention

[0087] Take 0.2M, pH7.3 HEPES-NaOH buffer, add TOOS, 4-aminoantipyrine, peroxidase, adenosine deaminase, xanthine oxidase in turn, and finally add HEPES-NaOH buffer to set The chromogenic solution contained TOOS at a final concentration of 40mM, 4-aminoantipyrine at 6g / L, peroxidase at 13U / mL, adenosine deaminase at 9U / mL, yellow Purine oxidase. It is the chromogenic solution of the present invention, which is placed in a brown bottle and stored at 4°C for future use.

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Abstract

The invention provides a method and a developing solution for determining ribosome inactivating protein activity and application of the developing solution. The method for determining the ribosome inactivating protein activity is simple to operate, low in cost and capable of completing a quantitative determination process only with a spectrophotometer, and can avoid interference of traditional RNA enzyme on reaction, thereby being promising in practical application prospect for the research and development field of ribosome inactivating protein.

Description

technical field [0001] The invention belongs to the technical field of biochemical analysis and detection, and in particular relates to a method for measuring the activity of ribosome inactivation protein. Background technique [0002] Ribosome-inactivating protein (RIP) is a kind of phytotoxic protein that can inactivate eukaryotic ribosome and inhibit protein synthesis, and widely exists in angiosperms, bacteria, fungi and algae. RIP has broad-spectrum anti-tumor, anti-virus, immune regulation and other biological activities, and has little toxicity to normal cells. It has received extensive attention and research all over the world in the past two decades. [0003] But there is an obvious problem in the development of RIP at this stage—there is no simple, convenient and reliable method to measure the activity of RIP. The existing methods roughly include the following: First, the activity of RIP is calibrated by measuring its inhibition of tumor cell proliferation in vitr...

Claims

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Application Information

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IPC IPC(8): C12Q1/34C12Q1/28C12Q1/26
CPCC12Q1/26C12Q1/28C12Q1/34C12Q2326/96
Inventor 孟尧孟延发
Owner CHENGDU MEDICAL COLLEGE
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