According to the statistics of "Nature", more than 70% of researchers have tried but failed to reproduce another scientist's experiments, and more than half have failed to reproduce their own experiments. One of the major factors contributing to the lack of reproducibility is the reagents used. Especially, the antibody reagents have an important role in the repeatability of publication data.
All of Sino Biological antibodies are developed and manufactured independently, and all the validation results are in the open. In order to provide customers more reliable antibodies, we have invested a lot of time and money in antibody quality verification and have established an highly strict QC process and various analytical methods.
The overall antibody validation steps in Sino Biological are shown as follows by taking the example of IHC antibodies.
|Antigen Design||→||Multiple Experimental
|→||Antigen Blocking||→||Multiple Tissues||→||Batch Stability|
|A: Blank control||B: Negative control||C: Positive control|
A: Blank control: There is no primary antibody in order to ensure that the secondary antibody does not non-specifically bind to certain cellular compartments.
B: Negative control: The protein you're interested is not present in the tissue. Therefore, if you see staining in this type of control, it indicates that the staining is non-specific.
C: Positive control: This is a tissue known to express the protein you are detecting. A negative result from the positive control demonstrates that the procedure has gone wrong.
Antigen blocking means a method on the basis of pre-incubation of primary antibody with the immunogen to confirm the specificity of antibody. If the tissue shows no staining with antigen blocking, it is a powerful evidence that the antibody is specific to the target you're interested. But there are special cases. For instance, when the immunogen is the whole protein, the mixture of antibody plus protein may result in higher non-specific staining, but the mechanism is unclear now.
|Multiple Tissue Samples|
|Tonsil sample 1||Tonsil sample 2||Tonsil sample 3|
|Prostate sample 1||Prostate sample 2||Prostate sample 3|
|Multiple Tissue Types|
Your target protein may express in different tissues, while usually the staining results of an antibody in different tissues are not exactly the same. Can you detect the target protein in the tissue you want with the antibody? This directly affects your experimental results. The test tissue provided by the antibody supplier may not be the same as the sample you want to study. Therefore, when the antibodies are validated in more tissue types and samples by the supplier, you can refer to more information and your experiments are more likely to be successful.
|Strict lot-to-lot validation to ensure the reproducibility|
|Batch 1||Batch 2||Batch 3|
|Batch 1||Batch 2||Batch 3|
|Validation of Antibody Stability|
|Newly produced||After one year||After two years|
Due to the inevitable differences existing in the animals immunized, different batches of polyclonal antibodies may not perform exactly the same in IHC staining. Although monoclonal antibodies are secreted by the same B lymphocytes, the changes in cell culture conditions and antibody purification methods may result in the changes in antibody properties. Therefore, whether it is the polyclonal antibody or the monoclonal antibody, the stability of antibodies between batches is of great importance.