Suppose if mRNA is showing up regulation of a gene through qRT-PCR, then, is it necessary that the level of corresponding protein will also be high in western blot.
Hi Sandeep,
It is likely that the amount of protein will be increased as well. However, there are several mechanisms which could prevent an increased protein generation e.g. increased degradation of the mRNA or inhibition of the translation by other RNAs.
Hi there,
there is no strict relation between mRNA level and protein level.
If simply you wanna check the protein transcription then off-course westernblot must have to indicate, as you mentioned.
Apart from this as @Boas indicated that are many factors involved. For instance, nuclear and cytoplasmic proteins will react differently because of the extraction methods. Some metabolic pathways can also affect the expression of your protein. In short you must have much more information about the behavior of your protein.
Not necessary, for example, I worked with heterologous protein expression in plants and their have several mechanism RNA silencing and therefore limits transient expression of heterologous proteins in plants. However, you can find several alternatives to improve silencing suppression depending on the expression system. If you are working with plants maybe this article can be useful: http://www.ncbi.nlm.nih.gov/pubmed/23933077, to give an idea.
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