Most of the literature available determining the expression of mTORC1 reports only western blot analysis. Could not find any PCR analysis
Well, what kind of information are you trying to get about mTORC1? RNA expression pattern in mouse tissues/cell lines or primers working for qPCR analysis ?
In general, protein expression is checked by immunological tests such as ELISA or western blotting or immunohistochemistry. RT-PCR would be used for transcript analysis, if you wish in parallel. But, if you wish PCR amplification of the antibody-mediated signal of protein expression, you can go for immuno-PCR!
We are looking for the expression of mTORC1 in the MCF-7 cells treated with anticancer compounds. Could not find primers for running PCR
If you access NCBI site to retrieve mTOR sequence from a species close to your cell line, you will be able to select your own PCR primers using Primer3.0 or any other suitable software. mTOR1 gene being conserved, the primers based on conserved regions of the sequence should work in your study on MCF-7 cell line. It is worth trying that way.
Hi, Shabana, I strongly recommend you go to Roche Universal Probe Library to design your own qPRC primer sets. It's a free and handy tool for designing qPCR primer. And the best thing is you don't need optimize PCR temperature because all the primers it generates have been optimized for annealing temperature 60oC.
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