According to the catalytic characteristics of HRP , ELISA Generally use hydrogen peroxide (H2O2) as one of the substrates HRP , When the hydrogen supply ( is the original substrate ) , the response of HRP and H2O2 is rapid and dedicated. From Figure 4 , it can be seen that HRP was oxidized by H2O2 dual -valent i , and the complex i ] It can also be restored to the starting state with the two -step unit price of the hydrogen supply. The complex Ⅱ is an intermediate product with an oxidized electron. When H2O2 is excessive, due to the formation of the complex Ⅲ or ⅳ, the enzyme activity is suppressed ( later ) . 30 % H2O2 is not stable. Because H2O2 is both a substrate that is HRP and it is also a inhibitor, Elisa To get a satisfactory measurement result, h2O2 must be necessary Limited to a certain concentration range, the final concentration is usually 2 ~ 6 mmol/l . However, in actual research work, we rarely pay attention to this. The concentration of H2O2 used by most researchers is often larger than the amount required for ideal reactions 2 ~ 4 . HRP adsorbed by solid phase is more vulnerable to overdose HRP . If 30 % H2O2 Storage concentration is confirmed to be 30 %, then dilute 10 000 - 12 000 [ 123] Double is the ideal substrate. H2O2 [123 ]'s Moore's lighting coefficient is 10mm light diameter 240nm is 43.6 . Therefore, it can be detected in this way [123 ] H2O2 Work solution concentration. Solid phase Elisa , when the temperature is higher than 20oc

,

HRP is often low, and non -ions are added to the substrate solution to the substrate solution Snueling agent polycanol -20 or TritonX-100 Can delay the loss of HRP and increase the reaction temperature, but the enzyme activity protection effect of non -ions deodorant is different according to the different supply of hydrogen.For example, 2,2 '- Lian Nitrogen - Double -[3- ethylene pyrarararine ]-6- Sulfate (ABTS) is a hydrogen supply, which can only protect 20 % enzyme activity, and neighboring fennel amine (ODA) [) [) [ODA) [)123] The protection of enzyme activity increased to 90 % when it was a hydrogen supply body. HRP The reason for so far in Elisa is the most widely used labeling enzyme, mainly because it is easy to extract and the price is relatively low; the other isThe role of stable nature, heat resistance and organic solvents, after couplet with antigen or antibodies, the activity is rarely lost.

Remarks: Product information may be optimized and upgraded.Please refer to the actual label information.