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

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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.