SDS-PAGE analysis of one step purification of GFP from E.coli BL21DE3 whole cell lysates using GFP-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) GFP-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 M: 10 µl See Blue Plus pre-stained protein marker (Thermo).
Lane 1: GFP-spiked lysate containing 21 µg of total protein.
Lane 2: 16 µl of polyol eluant from the affinity resin, from 108 µl total elution.