Profilin 1 (recombinant human no tag)

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Actin binding studies
Actin polymerization studies

Profilin is tested for biological activity by a binding assay with actin. Profilin is bound to poly-L-proline sepharose beads and incubated with G-actin. The profilin beads bind > 90% of the available actin by this procedure.
Actin Polymerization Inhibition Assay. The ability of profilin to inhibit actin polymerization was assessed by SDS-PAGE of proportionally loaded supernatant (S) and pellet (P) fractions from G-actin incubated with and without profilin-1 according to the assay method. In the absence of profilin-1, approx. 80% of the actin protein (43kDa) is found in the pellet fraction as F-actin (P, lane 2).When G-actin is incubated with profilin prior to polymerization, only 20% approx. of actin is found as F-actin in the pellet (P, lane 3), while the other 80% remains as G-actin in the supernatant (S, lane 3). Lane 1, profilin-1 protein alone. Mark12 molecular weight markers are from Invitrogen.