I currently work with tracheal MSCs and would like to measure the expression of aSMA.
I used bAct as housekeeper so far, but would like to use a different one for aSMA, because both proteins have the molecular weight.
I tried GAPDH, but I treat the MSCs with hyperoxia for 72h and it seams to have an effect on the expression of GAPDH.
Could you recommend a different housekeeper which works good for MSCs and has a different molecular weight then aSMA?
Thank you very much for your help.
Hi,
You can try beta-tubulin however it's of 50kDa which will appear almost in the region of aSMA. The best will be opting for stripping or you can load two sets of samples adjacently in one gel and run and stained separately. Alpha SMA molecular weight is 42kDa while the molecular weight of GAPDH, beta-actin, and beta-tubulin is 36kDa, 42kDa, and 50kDa resp. So, how to proceed will with western that is will matter, not which housekeeping or what is your cell type. alpha SMA expresses nicely in MSCs.
Regards
Saurabh
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