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 marker15 μl - IPTG culture sample15 μl + IPTG culture sample15 μl post-sonication pellet15 μl column flow through15 μl column wash through, 1st 10 ml15 μl column wash through, remainder15 μl fraction 315 μl fraction 415 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 marker15 μl fraction 615 μl fraction 715 μl fraction 815 μl fraction 915 μl fraction 1015 μl fraction 1115 μl fraction 1215 μl fraction 1315 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.

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