Nectagen

SDS-PAGE analysis of one step purification of a Thioredoxin-fusion protein from E. coli BL21(DE3) whole cell lysates using trxA-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) trxA-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: Thioredoxin-spiked lysate containing 21 µg of total protein.

Lane 2: 16 µl of polyol eluant from the affinity resin, from 127 µl total elution.