Figure 6

Using CPH substrates in simple agarose plate assays. (A) Schematic illustration of the assay using agarose plates. A well is formed in the agarose (either during or after casting), and CPH substrate is added together with an enzyme-containing solution. (B) Diagram showing the situation after the digestion period. If the enzyme has activity against the CPH substrate, then a halo is formed by the migration of soluble products. (C) and (D) examples of agarose plate assays with a glucanase (glc) used to digest four differently coloured version of CPH-pachyman (PACHY), and a xyloglucanase (xg) used to digest four differently coloured versions of CPH-xyloglucan (XYG), respectively. (E) An example of an agar plate assay containing mixed CPH substrates. All the wells contained red CPH-pachyman (PACHY), blue CPH-galactomannan (GALMAN) and yellow CPH-xylan (XYLAN) mixed in approximately equal proportions. Note that different coloured halos are generated depending on what enzyme or combinations of enzymes were present in wells and as indicated in the legend. See Tables 1 and 2 for details of the substrates and enzyme used.