Figure 1: Purification of HIV gag p24.
E. coli competent BL-21 containing His-gag p24 plasmid was grown overnight and induced by IPTG. The bacterial culture was spun down and the pellet lysed with extraction buffer (Histalon purification kit). The lysate was cleared by centrifugation and the supernatant was loaded into Histalon metal affinity column. Following washing, the column was loaded with elution buffer. The eluate was then collected, dialyzed against PBS, pH 7.4, and analyzed on 10% protein gel under reducing condition. Molecular weight marker was also included. The gel shows highly pure protein (gag p24) of approximately 78 KD in non-reduced form and 26 KD in the reduced form.