Elisa experiment optimizes the three conditions of the coating - Database & Sql Blog Articles

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First, how to choose the coating antigen

The coated antigen can be divided into three types: natural protein, recombinant protein and small molecule antigen. Natural proteins purify packaging requirements by direct adsorption, and many indirect methods of impurity antigens can capture data packets (the first and target antigen-reactive substances, such as direct adsorption to the ELISA plate, and then through the specific reaction of the solid phase of the antigen-antibody). Purification of recombinant proteins can usually be applied directly. Small molecule antigens such as peptides and some small molecule organic compounds, whose molecular weight is too small, are difficult to adsorb directly on the ELISA plate, usually the first and unrelated proteins such as bovine serum albumin, coupled, coupled to a solid phase carrier for adsorption.

Second, how to choose the coating liquid

Generally one

Ph9.6

Carbonate buffer, if the coating antigen is unstable under alkaline conditions, it can also be used

pH 7.2

Phosphate buffer solution.

Third, how to choose the temperature of the coating

Usually in

4-8

Degree

2

Hour overnight or

37

Under the conditions of the degree, we strongly recommend

4-8

The temperature range is beneficial to maintain the activity of the protein. select

4

The optimum concentration of the layer concentration coated solid phase carrier and coating performance change substance, total protein coating concentration

1-5ug

/

Ml

The experimental determination of the specific concentration of the specific antigen should be determined by the coating.

Whether it needs to be closed after coating

?

How should the closed system be selected?

?

Whether blocking is necessary depends on the enzyme-linked immunosorbent model and the specific experimental conditions. In general, the double antibody sandwich method can provide satisfactory results as long as the enzyme label is highly active, and the operation is thoroughly cleaned without closure. Determination and indirect methods, general closure is essential. Common sealant

0.05%

0.5%

Bovine serum albumin,

10%

Fetal bovine serum, gelatin

1%

Skim milk powder

5%

,

Abmart

Recommended

5%

Skim milk powder, cheap and sealing ability, but skimmed milk powder

5%

Only suitable for short-term use, do not store for a long time, so use fewer kits.