Microwaving for double indirect immunofluorescence with primary antibodies from the same species and for staining of mouse tissues with mouse monoclonal antibodies |
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| Authors: | D. Tornehave D. M. Hougaard L. -I. Larsson |
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| Affiliation: | (1) Department for Pathobiology and Diagnostics, Danish Veterinary Institute for Virus Research, Lindholm, Kalvehave, Denmark, DK;(2) Department of Clinical Biochemistry, Statens Serum Institut, Copenhagen, Denmark, DK;(3) Division of Cell Biology, Department of Anatomy and Physiology, The Royal Veterinary and Agricultural University, Gronnegaardsvej 7, 1870 Frederiksberg C, Denmark e-mail: lail@kvl.dk Tel.: +45-35282851, Fax: +45-35282547, DK |
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| Abstract: | Many methods have been devised for double immunocytochemical staining. We now describe that moderate microwaving does not elute antibodies, but prevents their reactions with subsequently applied reagents. Thus, microwaving performed in between the first and second staining cycles permits double indirect immunofluorescence staining with antibodies raised in the same species. Moreover, microwaving also inhibits reactions with endogenous immunoglobulins present in extracellular compartments. This substantially reduces background in indirect immunostaining of mouse tissues with mouse monoclonal antibodies. Accepted: 19 October 1999 |
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