Localisation of Thermomyces lanuginosus SSBP xylanase on polysulphone membranes using immunogold labelling and environmental scanning electron microscopy (ESEM)

摘要

Antibodies were raised against purified xylanase from Thermomyces lanuginosus SSBP in mice. The anti-xylanase antibody had a log_2 dilution of four at day 39 as determined using enzyme-linked immunosorbent assay. The xylanase enzyme was adsorbed onto polysulphone membranes and the primary antibody was attached. An immunogold labelled secondary anti-mouse antibody was attached to the primary antibody to locate the enzyme on the membrane. Using an environmental scanning electron microscope, immunogold labelled xylanase was visible at 50 000 x magnification on the surface and the pores of the membrane. Controls showed no sign of immunogold particles.