Matt Gethers/CRI, Thailand/Labwork/HmgR Purification Protocol Using Heparin/poly-ac 7.11.08

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SDS-PAGE to Assay Elution Fractions

Gel 1

Lane 1 Lane 2 Lane 3 Lane 4 Lane 5 Lane 6 Lane 7 Lane 8 Lane 9
5 μl Unstained protein marker10 μl BL21 lysate supernatant10 μl BL21 lysate pellet10 μl column flow through10 μl column wash through10 μl fraction 310 μl fraction 510 μl fraction 610 μl fraction 7

Gel 2

Lane 1 Lane 2 Lane 3 Lane 4 Lane 5 Lane 6 Lane 7 Lane 8 Lane 9
5 μl Unstained protein marker10 μl fraction 910 μl fraction 1110 μl fraction 1310 μl fraction 1410 μl fraction 1510 μl fraction 1610 μl fraction 1810 μl fraction 20

The supernatant, pellet, and flow through lanes were just messy blobs. Some smearing showed up in the rest of the lanes in gel 1, but nothing at the 28 kDa mark. All lanes in gel two had a band at 28 kDa with 14 and 16 being the strongest. I recall that when loading in lane 15, the well wasn't very cooperative and I didn't get much sample in. All lanes had some contamination above the the 28 kDa mark running between the topmost band of the marker and the 3rd from the top. There was also something right below the 5th band from the top in fractions 9 and 11. Fractions 13, 14, 15, 16, 18, and 19 looked pretty clean over all, though.

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