What are the points that we have to keep in mind while selecting a suitable loading control for tissue lysate and body fluid such as blood plasma? Please help me.
For tissue lysate you can use beta-actin or GAPDH and for blood plasma just load equal amount.
Depending on what is the target of your experiment, you should select loading controls among components that are not affected (at least based on our current knowledge) by the treatment or the condition you are studying. Usually, abundant proteins with a structural role (in the case of cells and tissues) are selected, such as actin or tubulin. Also, GAPDH or HSP70c can be used. Regarding blood plasma, albumin could serve as a loading control.
I agree with Amar Bahadur Singh, regarding the blood plasma loading control. The main problem with plasma is large variability in all protein concentrations in terms of subjects` health, nutrition and even the way of sampling. We used standard plasma as loading control and normalized Western blot results of target protein to it. The best you can do is to buy that standard plasma and load equal amount of your samples and standard plasma.
In fact using of GAPDH control in serum or plasma western blot samples isnt favourable, because this enzyme usually serve in the cytosol inside cells, and the amount of this enzyme in the circulating liquid is very small. So, I think the best choice is to use cmc or another control.
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