April 27, 2000 UCP antiboby test

The lanes contain human latissimus dorsii muscle (GDH), control yeast, and yeast expressing UCP1, UCP2, and UCP3. The top blot was treated with ant-UCP2 antibody, and the bottom blot with the anti-UCP3 antibody. The signal that appears with the GDH samples (at either end of the gel) are just above or at the 30 kDa marker (this signal is actually a doublet with the anti-UCP3; the bands can be better separated with different gel conditions).

With the UCP2 antibody, note the clear band in the UCP2 expressing yeast at about 32 kDa. The smear and other lower band at the bottom of this lane are, I believe, degradation products from overexpression. I do not know what the lower band recognized in the UCP3-expressing yeast is. I do not believe it is UCP3, because we don't see any band at 32-34 kDa where there should be at least some of the protein.

With the UCP3 antibody, no difference is observed between any of the yeast.