Comparison of immunological experiments

Immunology three tools, immunohistochemistry, Western, ELISA, for positioning, qualitative and quantitative.

IHC (Immunohistochemistry)

It is a chromogenic reagent (fluorescein) that combines immunological principles (antigen-antibody-specific binding) and histological techniques (tissue acquisition, fixation, embedding, sectioning, dewaxing, hydration, etc.) to chemically react to label antibodies. , enzymes, metal ions, isotopes) color, to locate, qualitative and quantitative study of antigens in tissues (cells) (mainly localization). The sample is a cell or tissue. To observe the result under the microscope, membrane positive, qualitative positive, and nuclear positive may occur.

ELISA (Enzyme linked immunosorbent assay)

Using immunological principles and chemical reaction color development, the samples to be tested are mostly serum, plasma, urine, cell or tissue culture supernatant, so no tissue embedding, sectioning and other techniques are used, which is related to immunohistochemistry. The main difference, the operation begins to bind the antigen or antibody to the surface of the solid support, so that the later formed antigen-antibody-enzyme-substrate complex is adhered to the carrier, which is the meaning of "adsorption".

WB (Western Blot)

SDS-PAGE is first performed, and then the separated protein sample is transferred to a solid phase carrier by an electrorotator, and then the antigen-antibody-label color is used to detect the sample, which can be used for qualitative and semi-quantitative.

For IHC, positioning is the biggest advantage of immunohistochemistry. The method can accurately locate the antigen in tissues and cells, and thus can simultaneously observe different antigens in the same tissue or cells, so that the combination of morphology and function can be studied, compared with other protein detection methods. Immunohistochemistry has direct and accurate localization and high sensitivity. It is a preferred method for localization detection and analysis. It is very meaningful to carry out in-depth research in the field of pathology, especially for translocation studies of some factors.

Compared with IHC technology, WB may be more accurate. Of course, WB can also be characterized and localized (by extracting membrane proteins or nuclear proteins, cytosolic proteins to detect the antigen content, respectively, and indirectly reflecting their localization), but the sensitivity is far Below IHC.

Compared with immunohistochemistry, ELISA is the most accurate and one of the preferred methods for the detection of secreted proteins. Especially useful for micro-quantitative analysis of multiple samples, the amount of reagents and samples used is small and the results are accurate. IHC is highly targeted and is typically sampled from tissue or cells, while ELISA typically uses serum or tissue milling. Immunohistochemistry can quickly detect positive or negative antibodies in the sample, and is generally not used for quantitative detection.

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