Figure 3
Characterization of crosslinked product for molecular exclusion. 2 mg of IN 1–105 were crosslinked in 200 μL and loaded on a Superdex S-200 column equilibrated in 10 mM tris pH 7.5, 0.5 M NaCl, 1 mM DTT and 5% glycerol at a flow rate of 0.5 mL/min. 0.5 mL of each fraction were collected. Protein elution was monitored by the absorbance at 220 nm. The chromatographic profile of the crosslinked protein is shown in black, and unmodified control protein is shown in red. The elution position of molecular weight markers is indicated by arrows. The markers used were: β-amylase (200 kDa), serum albumin (66 kDa), carbonic anhydrase (29 kDa) and cytocrome C (12.4 kDa). The (V0) indicated the void volume (dextran blue).