Matt Gethers/CRI, Thailand/Labwork/HmgR Purification Protocol Using Heparin/7.28.09 SDS-PAGE Results

SDS-PAGE to Assay 7.28.08 Elution Fractions

Gel 1

Lane 1	Lane 2	Lane 3	Lane 4	Lane 5	Lane 6	Lane 7	Lane 8	Lane 9	Lane 10
5 μl Unstained protein marker	15 μl - IPTG culture sample	15 μl + IPTG culture sample	15 μl post-sonication pellet	15 μl column flow through	15 μl column wash through, 1st 10 ml	15 μl column wash through, remainder	15 μl fraction 3	15 μl fraction 4	15 μl fraction 5

Gel 2

Lane 1	Lane 2	Lane 3	Lane 4	Lane 5	Lane 6	Lane 7	Lane 8	Lane 9	Lane 10
5 μl Unstained protein marker	15 μl fraction 6	15 μl fraction 7	15 μl fraction 8	15 μl fraction 9	15 μl fraction 10	15 μl fraction 11	15 μl fraction 12	15 μl fraction 13	15 μl fraction 14

I don't see any sign of an ~28 KDa protein on the gel. The -/+ IPTG samples from the culture look identical, so I don't think I'm getting any induction. There's plenty of protein in the post-sonication pellet and the flow through, a little bit in the first fraction of the wash through, and nothing much in the last fraction of wash through. In each case, there is protein along the length of the lane, no real peaks. Fractions 3 - 5 of the elution have several bands, again none of which are ~28 KDa. Fractions 6-14 also have several bands, though there appears to be a recurrent band at ~100 KDa.