The verification can be done from the following three aspects:
First, at the molecular sequence level, after constructing the expression vector, the expression plasmid is sequenced for confirmation. Sequencing files can be used for verification.
Second, during the expression process, a comparison is made between induced and uninduced lysis buffers. Differences in protein bands can be detected by SDS-PAGE. The difference can be observed at the theoretical size of the target protein.
Lastly, Western blot (WB) validation can be performed using tag antibodies or antibodies specific to the target protein.
If protein verification is needed, protein coverage analysis can be conducted by comparing the theoretical sequence with the actual coverage ratio. However, the purity of the target protein must be greater than 90%. If the purity is lower, the coverage ratio may be affected. For lower purity samples, gel slicing can be performed for further analysis.