Antibody, on the one hand, is the raw material for the research and development of immunodiagnostic reagents, on the other hand, is the object for the detection of immunodiagnostic reagents. It has always been the main force in the field of biological science, but its reliability has repeatedly put researchers in a dilemma. For a long time, there are unavoidable quality problems in antibody products. As a result of antibody quality, the global research fund loss is as high as 800 million US dollars, which directly accounts for more than one third of the total market value of the whole research antibody. In order to avoid the problem of quality antibody, the most effective way to improve the repeatability of the test is antibody verification.

Affinity conjugates / antibodies are one of the most commonly used tools in life science. They are used to study proteins and their functions in various biological pathways and diseases. The specificity and sensitivity of these tools can help people easily identify specific proteins at various levels of expression.

But more and more studies show that not all of these tools are specific, which brings increasing problems to the repeatability of experiments, resulting in waste of time and resources and even recall of articles in some cases.

Antibody is one of the most commonly used tools in the field of life science. WB is the most commonly used method to detect a protein in complex samples and analyze it quantitatively. In addition, they are also used for immunohistochemistry and immunofluorescence staining to visually observe proteins under microscope, as well as other kinds of experiments based on the binding of antibodies to specific biomolecules.

It is much more complicated to verify antibody than to verify cell. Whether the antibody can work correctly depends on the specific test. For example, some antibodies can detect denatured proteins, but not denatured proteins, or vice versa. Antibodies that work well in one tissue may give false signals in other tissues or backgrounds. We also emphasize that antibody verification should be carried out according to its application and background.

Knockout is one of the most accepted and trusted methods to verify antibody specificity.

In a knockout cell line, a protein target does not exist because the gene expressing a specific target protein is removed, also known as knockout. The results will be compared with the control cell line (or wild type). If the antibody is truly specific, the results will not be detected in the knockout cell line, but specific signals will be detected in the control cell line. Knockout validation provides a completely negative control group to confirm the specificity of the antibody to a specific target protein.

Example of knock-out antibody verification - Caspase3 multi antibody specific knock-out verification: