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Figure 1

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NusA purification by nanoCLAMP nusA-A1 affinity chromatography

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SDS-PAGE analysis of one step purification of NusA from E.coli BL21DE3 whole cell lysates using nusA-A1(Resin) affinity resin. E.coli were lysed with BPER (Thermo), cleared by centrifugation, and diluted with PBS to 1.2 mg total protein/ml. The lysate was spiked with recombinant antigen to 0.1 mg/ml in 1.4 ml and incubated with 10 µl (packed vol) nusA-A1(Resin). Beads were washed with PBS and eluted with polyol elution buffer at near neutral pH. Lysates and eluants were analyzed on 12% SDS-PAGE in reducing SDS sample buffer.

Lane 1: NusA-spiked lysate containing 10 ug total protein.

Lane 2: 6.5 µl of polyol eluant from the affinity resin, from a total of 125 µl total elution. Note: the band in lane 2 at around 40 kDa was analyzed by mass spec and found to be a fragment of NusA.