Preparation method of O-substrate phenylene diamine of horseradish peroxidase

Abstract

The invention discloses a method for preparing ortho-phenylene diamine substrate of horseradish peroxidase: firstly dissolving 100 mg ortho- phenylene diamine in 100 ml N,N-dimethyl formylamine to form solution A, where the oxydo-containing citric acid-phosphate substrate buffering solution is solution B, and if use, mixing 5 ml solution A and 10 ml solution B to make the ortho-phenylene diamine substrate. The invention mainly improves the formula of the ortho-phenylene diamine substrate of horseradish peroxidase, solving the problems that accurately weighing ortho-phenyene is complex, and that the ortho-phenylene diamine is uneasy to dissolve in the substrate buffering solution, and so on; besides, this improvement can not destroy the sensitivity of original horseradish peroxidase substrate system, nor destroy the analysis and testing of the system, and other related contents.

Description

technical field [0001] The invention relates to the field of immunoassay, in particular to a preparation method of o-phenylenediamine substrate of horseradish peroxidase. Background technique [0002] Horseradish peroxidase is a very important tool enzyme in the field of immunoassay. Due to its convenient source, low price, and sensitive detection system, it is widely used in immunological analysis and testing, among which there are soluble systems , Precipitating system, soluble system is that the final product is soluble after the substrate is catalyzed by horseradish peroxidase, so the data can be read by the instrument, so it is mainly used for quantitative and qualitative analysis. There are many substrates for horseradish peroxidase, the most commonly used of which is o-phenylenediamine, and the commonly used formula for this substrate is: [0003] 0.1M citric acid 24.3ml [0004] 0.2M Na2HPO4 25.7ml [0005] o-Phenylenediamine 20mg [0006] 30% hydrogen peroxide 7...

AI Technical Summary

Problems solved by technology

[0007] Although the price of o-phenylenediamine in the above-mentioned substrate formula is relatively low, the dosage of o-phenylenediamine is 20mg and it is difficult to accurately weigh the dosage in many units. The substance must be ready-to-use and ready-to-use, and it is sensitive to light. At the same time, this substrate is not easily dissolved by the substrate buffer when it is prepared, which brings inconvenience to laboratory research analysis and analysis and application of low-level analysis institutions. At the same time, due to the fact that the actual application is not constant, sometimes people only need to use 4ml and sometimes only 10ml of substrate in the analysis and test, so the corresponding substrate also needs to be changed, and the weighing will be accurate. Indeed, it has become cumbersome, so that some institutions have to weigh extensively

The amount of o-phenylenediamine weighed is too much, which is also a kind of waste, and it will also have an adverse impact on environmental protection