PMID: 9438069Jan 23, 1998Paper

Blot analyses and immunocytochemistry of neural antigens with digoxigenylated primary and secondary antibodies

Brain Research. Brain Research Protocols
W HärtigV Bigl

Abstract

While the digoxigenin-anti-digoxigenin (DIG) method is currently the preferred tool for non-radioactive in situ hybridization this study extends its application field to Western blotting of proteins and summarizes advantageous properties of digoxigenylated antibodies in immunocytochemistry. An established protocol for the preparation of digoxigenylated primary antibodies is complemented by dot blot analyses confirming the high sensitivity of hapten-anti-hapten techniques based on primary digoxigenylated antibodies. The comparative Western blot analysis of calcium-binding proteins in nervous tissue is used as an example to show the highly specific detection of relevant antigens with unmodified primary antibodies, digoxigenylated secondary antibodies and anti-digoxigenin-peroxidase conjugates. The application of the DIG technology seems to be especially indicated in tissues containing high amounts of endogenous biotin-bearing proteins which might induce false-positive staining in conventional streptavidin/biotin techniques. Finally, the previously shown suitability of digoxigenylated antibodies for different immunocytochemical procedures is completed here by examples for sensitive single immunoperoxidase staining of neural marker...Continue Reading

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Citations

May 5, 2001·International Journal of Developmental Neuroscience : the Official Journal of the International Society for Developmental Neuroscience·E KirazovR Schliebs
Mar 17, 2006·Journal of Neurochemistry·Katrin NowakSteffen Rossner
Dec 11, 1999·Circulation Research·J H CampbellG R Campbell
Jun 2, 2006·Neuroreport·Wolfgang HärtigRalf Hoffmann

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