Question (2036)
Question
I used abm’s virus to transduce my cell line. I determined that there is mRNA expression of my GOI but no protein expression. Why?
ABM community
Verified customer
Asked on Mar 24 2025
Answer
There could several reasons for this:
1. qPCR detection is much more sensitive than Western blot, and any target signal is amplified greatly during qPCR.
2. Verify that your primary antibody is of good quality and include good positive and negative controls in your Western blot.
3. Western blot can only be performed after a stable cell line has been established and notably it rarely works well with a polyclonal cell culture.
4. mRNA expression does not always correlate with protein expression because many different biological factors may affect translation, including protein half-life, protein degradation and molecular processes such as phosphorylation, ubiquitination, methylation, etc. abm is not in a position to guarantee GOI expression at the protein level due to the number of experimental variables involved. abm guarantees GOI expression at the mRNA level only, while expression at the protein level must be determined experimentally. We recommend checking the literature to see whether other scientists have been able to over-express the protein in the same target cells and how this was achieved (tag, delivery system, detection method, etc).
1. qPCR detection is much more sensitive than Western blot, and any target signal is amplified greatly during qPCR.
2. Verify that your primary antibody is of good quality and include good positive and negative controls in your Western blot.
3. Western blot can only be performed after a stable cell line has been established and notably it rarely works well with a polyclonal cell culture.
4. mRNA expression does not always correlate with protein expression because many different biological factors may affect translation, including protein half-life, protein degradation and molecular processes such as phosphorylation, ubiquitination, methylation, etc. abm is not in a position to guarantee GOI expression at the protein level due to the number of experimental variables involved. abm guarantees GOI expression at the mRNA level only, while expression at the protein level must be determined experimentally. We recommend checking the literature to see whether other scientists have been able to over-express the protein in the same target cells and how this was achieved (tag, delivery system, detection method, etc).
ABM Scientific Support
Answered on Mar 24 2025