I noticed a drastic expression difference for the same protein between Arabidopsis and rice. I am wondering why. Though I used equal amount plant material for immunoblotting, my protein abundance could be different.
Therefore, I am wondering how much proteins difference between the two plants? I checked online that Arabidopsis has 35,000 proteins, but I cannot find the information for rice.
Would RT-PCR be a better idea to check expression?
Thank you!
So you mean number of proteins in each plant? I would suggest you to measure the protein amount using BCA, Bradford or other assays. Than load the same amount for comparison. This is for common western blot. But I am not sure whether it is applicable for the problem you want to solve.
RT-PCR detect the expression at RNA level, which can't reflect regulation on productivity of translation. It can be useful if the differential expression is due to RNA change.
When doing these comparisons, first, load similar amount of protein or RNA; second, choose proper internal reference. These are to exclude the possibility that the observed differences are merely due to loading differences or artificial factors.
This paper describe the estimated number of protein-coding genes in rice genome
http://www.nature.com/nature/journal/v436/n7052/full/nature03895.html
However, no one knows the numbers of proteins because one gene might produce different protein isoforms. When it comes to a cell, the question is even more difficult because different cell types have different set of genes actively expressed and different number of proteins are produced in that cell type. So, the better way is to determine what type of cell you are interested in.
Thank you so much for the answer and the paper recommended. I would try equal protein loading and have a control protein antibody next time.
Yuan
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