Why is the density of L-type (Ca1.2) alpha subunit so low in HEK cells (assayed by whole cell voltage clamp)?
Have you tried co-transfecting the alpha1 subunit with the other ancillary subunits (beta, and alpha2 delta) or at least just the beta subunit? I found co-expression of ancillary subunits (or just beta) improves expression quite a bit. Best of luck!
Thanks Joyce, i should have mentioned we contransfect with beta2 and still can only get about 150 pA of current with Ca++. With Ba++ 15 mM its better, but i would like to get a couple of nanoamps.
Dear Arthur, I believe that final IBa was about 30-50 pa/pf which is fine for HEK cells, taking into account their low capacitance. There is paper, which possibly help you http://ajpheart.physiology.org/content/278/1/H126.long. B2a subunit also increases current itself, so this co-transfection must work. Good luck!
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