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Do you know about western blotting?

2021-08-18
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IBT and LIA

The Western blot detection technology currently used is not the original Western blot (IBT) but an optimized and improved method.

western blotting
western blotting

Traditional Western Blotting (IBT): the protein is separated and purified by electrophoresis and transferred to a nitrocellulose membrane, and then autoimmune serum is used as a probe to react with and bind to the antigen on the membrane. The antibody is detected by the color reaction between the enzyme-linked immunoglobulin antibody and the substrate. Although this method combines the high resolution of polyacrylamide gel electrophoresis and the high sensitivity of biomolecular affinity technology, it has many shortcomings. 1) Although the molecular weight of the traditional IBT method is different in the electrophoretic separation of proteins, because of their different charges, the swimming speed in the electric field can still be the same, so that the proteins of different molecular weights may have bands with the same electrophoretic distance. 2) The slope of the protein bands during electrophoresis brings certain difficulties to the identification of these bands. 3) The antigen in the traditional IBT method is denatured. Even though some antibodies may recognize the denatured epitope on the transfer membrane, not all antibodies can react with the denatured epitope. 4) The traditional IBT method cannot detect antibodies against conformation-dependent epitopes, resulting in missed detection of certain antibodies. For example, the epitope conformation of SSA autoantigen in autoantibody repertoire detection will be destroyed by gel electrophoresis.

Linear immunoassay method (LIA): It can also be called the enzyme immunobanding method. This technique is to coat the antigen purified by affinity chromatography or the antigen purified by genetic recombination technology on a fixed nitrocellulose membrane Fixed position. Each antibody corresponds to only one band, no non-specific reaction bands are generated, and can be customized according to the needs of disease detection. This new detection method has a high degree of specificity and sensitivity, and the results are accurate and easy to interpret. At present, Yahuilong’s blotting detection platform has:

Multiple indicators: provide the most comprehensive and reasonable combination of indicators, up to 17 indicators can be tested in one experiment;

Project full: antinuclear antibody spectrum, vasculitis spectrum, autoimmune liver disease spectrum, autoimmune diabetes spectrum, autoantibody comprehensive spectrum, etc. to meet different clinical needs;

Easy to interpret: use linear immunoassay method, and each/two indicators correspond to a membrane block, the color is clear and easy to interpret;

Accurate results: The antigens are all highly purified or recombined, containing weak positive, medium positive, and strong positive quality control bands. The interpretation is accurate and reliable;

Simple operation: supporting Tenfly Auto automatic western blotting analyzer automatic operation, automatic interpretation, providing a perfect overall solution for western blotting.

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Western Blot Analysis Services

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