The conjugate, an enzyme-labeled antibody (or antigen), is a key reagent in Zui in ELISA. A good conjugate should be to retain both the catalytic activity of the enzyme and the immunological activity of the antibody (or antigen). There is a suitable molecular ratio between the enzyme and the antibody (or antigen) in the conjugate, and the binding reagent should contain as little or no free (unbound) enzyme or free antibody (or antigen) as possible. In addition, the combination still has good stability.
The enzyme used in the ELISA should meet the following requirements: high purity, high conversion rate of catalytic reaction, strong specificity, stable nature, abundant source, and inexpensive price. After preparation of the enzyme conjugate, it still retains its active part and Catalytic ability. Zui is good, the same enzyme does not exist in the specimen. In addition, its corresponding substrate is easy to prepare and store, and the price is low, and the colored product is easy to measure.
In ELISA, commonly used enzymes are horseradish peroxidase (HRP) and alkaline phosphatase (AP). Among the few commercial ELISA reagents, the enzymes used are glucose oxidase, β-D-galactosidase and urease.
Domestic ELISA reagents generally use HRP to prepare conjugates. HRP is a glycoprotein with a sugar content of about 18% and a molecular weight of 44,000. It is a complex enzyme composed of a main enzyme (enzyme protein) and a prosthetic group (heme). It is a porphyrin. protein. The main enzyme colorless glycoprotein has a high absorption peak of zoi at a wavelength of 275 nm, and the prosthetic group is a dark brown iron-containing porphyrin ring having a high absorbance peak at 403 nm. The purity of HRP is represented by RZ (Reinheit Zahl, German, meaning purity), which is the ratio of the absorbance at 403 nm to the absorbance at 280 nm, and the RZ of high purity HRP is ≥ 7.0.
In addition to the above-mentioned requirements for labeling enzymes in ELISA, HRP is more inexpensive and more stable. It is worth noting that in the selection of enzyme preparations, in addition to its purity RZ, more attention should be paid to the activity of the enzyme. High-purity enzymes, such as improper preservation, will also reduce their vitality. The viability of the enzyme preparation is expressed in units of enzyme activity contained, and can be tested by measuring the amount of product produced by the action of the substrate.
Many ELISA reagents abroad use alkaline phosphatase (AP) as a labeling enzyme. Commonly used APs come from two sources, extracted from E. coli and calf intestinal membranes. The biochemical characteristics of the enzymes from different sources are slightly different. The molecular weight of AP extracted from E. coli is 80,000, the pH of the enzyme-treated zui is 8.0, the molecular weight of AP extracted from the calf intestinal membrane is 100,000, and the pH of Zui is 9.6. In the ELISA, the sensitivity of the AP system is generally higher than that of the HRP system, and the blank value is also low, but the AP is expensive, and the yield of the prepared conjugate is also lower than that of the HRP.
Antigens and antibodies The antibodies used in the preparation of conjugates are generally higher purity IgGs to avoid interference with other heteroproteins when linked to enzymes. Zui uses affinity-purified pure antibodies, so that all enzyme conjugates have specific immunological activities, and can be reacted at high dilution. The results of the experiment are light. If labeled with F(ab)2, RF interference in the specimen can be avoided. There are not many patterns of enzyme-labeled antigens in ELISA, and the general requirement is that the antigen must be of high purity.
A dilution of the conjugate is used to dilute a high concentration of the conjugate to formulate a working fluid. In order to avoid direct adsorption of the conjugate to the solid support in the reaction, a high concentration of unrelated protein (for example, 1% bovine serum albumin) is often added to the dilution buffer to compete for inhibition of conjugate adsorption. A nonionic surfactant having an inhibitory protein adsorbed on the surface of the plastic is also generally added, such as Tween 20, and a concentration of 0.05% is suitable. When the antibody is indirectly measured, the serum sample is diluted and measured, and the dilution can also be applied.
Introduction:
Hydrogen-rich water cup, also known as water-element water cup, originated in Japan. According to Japanese medical research, active small molecule hydrogen can provide comprehensive anti-oxidant care for the body, specifically for metabolic function repair, immune regulation, eliminating inflammation, improving allergic constitution, preventing cell mutation (cancer prevention and anticancer), promoting tissue repair, anti-aging beauty and beauty.
At the same time, hydrogen is easier to dissolve in water after decomposition. We know that hydrogen is usually insoluble in water. Therefore, only after molecular refinement, can it be integrated into water to a certain extent and achieve the role of increasing the content of hydrogen molecule.
The principle of hydrogen rich water:
Bacteria and viruses invade the body constantly through the food and breathing, However the white blood cells can release reactive oxygen species to protect us. Reactive oxygen will attack human normal cells and DNA, The components of cells will be oxidized, This is the cause of disease, aging and spot.
Hydrogen rich water contains rich hydrogen molecules. it will be decomposed into hydrogen atoms after drinking. With the human body`s reactive oxygen species, oxygen radicals undergo a reduction reaction to generate the most harmless water that the human body needs most. The chemical reaction with oxygen is called oxidation reaction, and the chemical reaction with hydrogen is called reduction reaction. The research report confirms that hydrogen is colorless, odorless, non-toxic, and has no side effects.
Hydrogen is one of the best natural antioxidants, because the hydrogen molecules in the hydrogen-rich water are very subtle, can penetrate cells quickly, neutralize the human body`s [health killer" oxygen free radicals, and finally generate water to be excreted. Hydrogen is hardly soluble in water, but the use of nano-level technology can solve this technical problem. Each hydrogen atom will uniformly wrap water molecules, thus forming small molecules, which can pass through cell membranes because of their small size, to liminates malignant oxygen free radicals in cells.
Why can hydrogen water improve physical health?
As we all know, the human body is composed of cells, the human disease can eventually be attributed to the cell by Loss of human aging is due to cell aging or necrosis caused by. And cause cell disease sick old The main culprit is the excess oxygen free radicals. The main effects of oxygen are: antioxidant. A hydrogen atom Combined with reactive oxygen species, reduced to water, excreted.
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