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An evaluation of the conditions necessary for optimal protein a gold labeling of capsular antigen in ultra thin methacrylate sections of the bacterium pasteurella haemolytica

, : An evaluation of the conditions necessary for optimal protein a gold labeling of capsular antigen in ultra thin methacrylate sections of the bacterium pasteurella haemolytica. Histochemical Journal 16(2): 151-164

The protein A-Au (PAG) probe is a particulate immunocytochemical probe that is eminently suitable for quantification. The effect of probe concentration, variation of labeling on different sections within a single grid, washing procedures, variation of labeling with time and temperature and different storage conditions on the probe were investigated using PAG labeling of capsular antigen on ultrathin methacrylate sections of the bacterium P. haemolytica. In this antigen-antibody system, and using a 20 nm probe, optimal results were achieved with 2 .times. 1012 particles/ml, a labeling time of 60 min at room temperature; the PAG probe, stored at C, should be 1-5 wk old. The efficiency of the probe tested by evaluating different primary antibody concentrations, by evaluating cross reactions of the primary antibody and by evaluating the relative amounts of antibody against internal components of the bacterium present in different antisera.


PMID: 6698803

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