Question (2040)
Question
I performed gene knockout using abm’s CRISPR knockout vector/virus. The western blot revealed that my protein is still present. Why?
ABM community
Verified customer
Asked on Mar 24 2025
Answer
There could be several possible reasons for this:
1. Cells are polyclonal. Cells are a mixture of clones, not a single clone. We recommend isolating single cells by serial dilution and then screening and sequencing and testing those cells.
2. The cell line may not be diploid, but instead could be triploid or more. In other words, the cells have more than two copies of the same gene. This is a very common phenomenon in cancer cell lines.
3. The antibody being used may be binding to other proteins from the same family. Assess the antibody and ensure that it only binds to your target protein.
1. Cells are polyclonal. Cells are a mixture of clones, not a single clone. We recommend isolating single cells by serial dilution and then screening and sequencing and testing those cells.
2. The cell line may not be diploid, but instead could be triploid or more. In other words, the cells have more than two copies of the same gene. This is a very common phenomenon in cancer cell lines.
3. The antibody being used may be binding to other proteins from the same family. Assess the antibody and ensure that it only binds to your target protein.
ABM Scientific Support
Answered on Mar 24 2025