Hello everyone, I would like to ask how to optimize the Western blot experiment for TMS1/ACS Antibody (HY-P80548) and Caspase-1 Antibody (HY-P81232), as the bands are very faint and there are non-specific bands.
Hi，
The following optimization steps can be taken:
1. Using blocking buffer with primary and secondary antibodies, or 0.5% BSA in TBST, can reduce background.
2. Adding 20% methanol to the transfer buffer can help transfer proteins from the gel to the membrane, improving transfer efficiency.
3. Increase the amount of protein loaded.
The answer to this question is provided by MedChemExpress Technical Support.
You can check the buffer conditions first. Run your protein of interest along with a positive control. Then cut the skim-milk–blocked membrane for your positive control and proceed with the remaining experiment using the same buffer. If your positive control works well, it means your conditions and buffer are fine. Next, optimize the dilution of your antibody by testing different dilutions. If you still encounter problems, it is better to try an antibody from another company.
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