The Applications Of The Antigen-antibody (P24Polyclonal And Monoclonal Antibody) Binding In Paraffin Tissue Sections

Objective:To verify the antigen-antibody (P24polyclonal and monoclonalantibody) binding in paraffin tissue sections and explore the workconditions.Methods:40Sprague-Dawley rats born in24h were intracranially (i.c.)inoculated in the right cerebral hemisphere with either BDV solution orphosphate-buffered saline (control: sham inoculated).40Sprague-Dawleyrats and40C57mice without injection were chosen as negative control.The animal brains were perfused at45days after birth, and weresubsequently embedded into paraffin. Then the immunohistochemicalstaining was performed follow Envision two-step protocol. The primaryantibody was chosen as P24polyclonal and monoclonal antibody and the dilution ranged from1:50to1:5000. The primary antibody was replaced byPBS as a blank control. We considered that brownish yellow or brownparticles distributed in the cells as positive, and scored with proportion ofpositive cells multiplied by the positive intensity score.Results:There is a significant difference in staining scores between theexperimental group and the control group (polyclonal P24group Z=-3.108,P=0.000, monoclonal P24group Z=-4.605, P=0.000. It has shown a highsensitivity with slightly stained background in dilution1:200,1:400usingpolyclonal P24staining while a high sensitivity without stained backgroundin dilution1:100,1:200,1:400using monoclonal P24staining.Conclusion:Our study suggested that BDV P24polyclonal and monoclonalantibody can bind the antigen distributed in paraffin sections of BDVinfectious rat. The recommended dilution of polyclonal P24antibody is1:200and1:400. The recommended dilution of monoclonal P24antibody is1:100,1:200and1:400.