ISHS Acta Horticulturae 110: V International Symposium on Virus diseases of Ornamental Plants
CONDITIONS FOR THE USE OF PROTEIN A IN COMBINATION WITH THE DERRICK METHOD OF IMMUNO ELECTRON MICROSCOPY |
| Authors: | D.-E. Lesemann, H.-L. Paul |
| Abstract:
The effect of protein A when used with the Derrick method of immuno electronmicroscopy, is characterized in respect of concentrations of protein A, antisera, and viruses. Protein A in a concentration of 10/ug/ml, used for pretreatment of grids prior to the coating with antiserum, were sufficient for obtaining high particle counts, which did not increase with more concentrated protein A. In contrast to the normal Derrick technique, protein A allowed to use concentrated sera which led to a high particle binding on antiserum coated grids, and to low unspecific particle adsorption on normal serum coated grids. Weak heterologous reactions could clearly be detected with the Derrick system when protein A was used. The characteristic saturation curve of particle numbers on antiserum coated grids in relation to the concentration of virus was only slightly altered by the use of protein A. Protein A did not lead to increased particle binding with dilute antisera or viruses, respectively. Thus the sensitivity of the Derrick system was not increased by protein A. | |
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