Antibodies are the most commonly used tools in biological research and are widely used in research techniques related to immunoassays, including enzyme-linked immunosorbent assay, western blotting, immunoprecipitation, and immunohistochemistry. Antibodies are also important tools for clinical monitoring and can be used for related diagnosis and treatment.
In recent years, scientists have found there has been a serious "duplication crisis" in antibody-related research. Antibodies used in studies vary widely in quality and consistency of performance, with significant adverse effects on the reproducibility of biomedical research.
To solve the antibody duplication crisis encountered in the market, ABclonal selects appropriate validation strategies based on the background data survey of the target, expression abundance, spatiotemporal expression specificity, and antibody detection threshold to ensure the antibody is correctly obtained and has the expected characterization. The validation strategy is as follows:
Genetic Strategies
In control cell/tissue samples, gene editing is applied to knock out or knock down the target gene, and when the expression of the target protein is eliminated or reduced by detection, the specificity of the antibody is determined.
Orthogonal Strategies
The target protein was detected in different samples by antibody-independent methods and compared to antibody-dependent detection methods. If the results are consistent, it indicates specificity between the antibody and the target protein.
Independent antibody strategies
Ensure two antibodies can bind to different regions of the antigen and recognize different epitopes, reducing the possibility of off-target effects.
Expression of tagged protein
Use affinity tags such as DDDDK, His, or fluorescent conjugated proteins such as GFP for detection. If the detection results of the anti-affinity tag antibody/fluorescence signal are consistent with the results of the antibody to be tested, the specificity of the antibody is confirmed.
Biological Strategies
Use biological methods to demonstrate antibody specificity for target proteins, such as comparisons across multiple cell lines (including positive and negative expressing cells) and multiple species.
Cell Biology & Developmental Biology
Protein phosphorylation
