Research elites! Note: 岚派生物 for your analysis of the advantages and disadvantages of the kit ELISA four methods - Database & Sql Blog Articles

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In immunoassays for the determination of proteins, antibodies, or hormones in liquid samples, ELISA assays are a broad assay technique. Research elites know that there are four methods commonly used in ELISA testing, so what are their advantages and disadvantages? Let's take a look at it with the e派生物elisa kit manufacturer Xiaobian.

1, direct method (direct ELISA)

The specificity of the primary antibody is very important by directly immobilizing the antigen on a solid phase carrier and adding an enzyme-labeled primary antibody.

Advantages: Short operating procedures, as there is no need to use secondary antibodies to avoid interaction.

Disadvantages: The primary antibodies in the test have to be labeled with enzymes, but not every antibody is suitable for labeling, and the cost is relatively high.

2. Indirect method (indirect ELISA)

This assay is similar to the direct method except that the primary antibody is not enzymatically labeled and the enzyme-labeled secondary antibody is used to identify the primary antibody to determine the amount of antigen.

Advantages: The secondary antibody can enhance the signal, and there are many options for different measurement and analysis. A primary antibody that is not labeled with an enzyme retains its most immunoreactivity.

Disadvantages: The probability of an interaction occurring is high.

3, double antibody sandwich method (sandwich ELISA)

The antigen to be detected is coated between two antibodies, one of which immobilizes the antigen on a solid support, ie, captures the antibody. The other is to detect the antibody, which can be directly labeled with the amount of the antigen after labeling with the enzyme; or without labeling, and then by the enzyme-labeled secondary antibody to determine the amount of the antigen. These two antibodies must be carefully selected to avoid interaction or to compete for the same antigen binding site.

Advantages: Highly sensitive, highly specific, the antigen does not need to be purified beforehand.

Disadvantages: The antigen must have more than two antibody binding sites.

4, competition method (competitive ELISA)

The antigen (free antigen) in the sample and the antigen (fixed antigen) purified and immobilized on the solid phase carrier compete for the same antibody. When the free antigen in the sample is more, the more antibodies can be bound, and the antigen is immobilized. Only bind to fewer antibodies and vice versa. After the washing step, the complex of the free antigen and the antibody is washed away, leaving only the complex of the immobilized antigen and the antibody, and compared with the result of the control group with only the immobilized antigen, the antigen in the sample can be calculated according to the color difference. content.

Advantages: Applicable to less pure samples, and the data reproducibility is very high.

Disadvantages: The overall sensitivity and specificity are poor.

The above are the advantages and disadvantages of the four methods of elisa detection. I believe that all the research elites are well aware of the chest. In the experiment, you can clearly choose the method that is most suitable for your own experiment, and match it with Shanghai Yipai Bio-elisa kit. The manufacturer's high quality ELISA kit will help you succeed!