For RNA-seq, what level of gene expression in TPM would be considered as expressed and can be detected by IF or FACS? I've seen a situation in which the level of CD68 is about 50(TPM) detected by RNA-seq, but IF and FACS are negative.
Its detected by RNA-seq exact method more than IF and FACS methods especially for small RNA amounts
Hey,
there might be another problem with what you are measuring, because having an expression on RNA level does not always mean you have a functional protein or an"expression" on protein level. There might still be something you are not considering, leading to an expression(TPM) but not a protein, meaning you do not have a signal in FACS.
Hope this helps a bit.
Regards,
André
@Yang, I agree with @Andre Marqurdt. Post translational modifications are very frequent and result in discordance between RNA expression and protein expression. If you are interested only in validation of RNA expression, real time PCR is a better option. If protein expression is what you want to measure, try western blot. There is just a little bit of chance that it may show something though IF and Flow are negative.
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