| Popis: |
In an attempt to determine the distribution of soluble complexes formed when 131I-labelled HSA reacts with fluorescein labelled anti-HSA we have used zonal centrifugation in A sucrose gradient combined with a subsequent analysis of the cuts by radioactivity, fluorescence and polarization of fluorescence measurements. The method is both reproducible and sensitive, and has a potential advantage over other methods for separating soluble complexes in that the sedimentation coefficients as well as antibody ratio of the complexes can be determined on microgram quantities of sample. Using this method, we have been able to follow the dissociation of antigen-antibody complexes with time, and to demonstrate the building up of large numbers of complexes at the equivalence zone. Full technical details of the method are given, together with suggestions for further improvements. |
