SDS-PAGE

Make up the gel according to instructions.

If you need to make more 10% APS, it's 0.1g APS per 900ul H20.

Cell pellets: use 1ml culture spun down at 13000g for 10 mins, discard supernatant and resuspend with 100ul sample loading buffer. Load 10ul on gel.

Amounts: 5ug - 10ug total protein.

5x sample loading buffer (Qiagen receipe):
0.225M Tris base pH 6.8, 50% glycerol, 5% SDS, 0.05% bromophenol blue.

To make 50ml:
11.25mL Tris base 1M
25ml glycerol
2.5g SDS Care: irritant
0.025g bromophenol blue

pH to 6.8

Mix 1:7 SLB:BME just prior to mixing with sample.