Silver Stained Tris-Tricine PAGE

Lane	Contents	Volume
1	Protein Molecular Weight Marker	20μL
2	Cbf5-Nop10-Gar1	20μL
3	Buffer A wash	30μL
4	Buffer B wash	30μL
5	Buffer E Wash	30μL
6	S-30 s/N	30μL
7	Buffer E – Concentrated Preprooxyphysin	30μL

To detect our prepro-oxyphysin proteins we did a Silver Stain 16.5% Tris-Tricine PAGE. The reason for doing a Silver Stain is that it is 10x more sensitive than the standard Coomassie stain. The tricine in the Tris-TRicine PAGE allows for better and more distinct separation of small proteins in the gel. In Lane 2, we did a control of Cbf5-Nop10-Gar1 proteins. Cbf5 is 43 kDa, Gar1 is 12.3 kDa and Nop10 is only 7.2 kDa. Nop 10 is outlined in red. Prepro-oxyphysin is slightly larger than Nop10 at 10kDa. The bands we suspect are prepro-oxyphysin are outlined by orange.