A new efficient immunoprotocol to detect chromosomal/nuclear proteins along with repetitive DNA in squash preparations of formalin-fixed, long-stored root tips

As a novel approach, the protocol includes an array of steps and options not described in chromosomal immunoprotocols that used aldehyde-fixed root tips for squashing, e.g., fixation with neutral-buffered formalin, storage of root tips in ethanol, squash in acetic acid, MWAR, protein redetection, immunoFISH-aided simultaneous DNA-protein visualization. It ensures chromosomal/nuclear spread of exceptional quality, rapid preparation of the fixing solution, prolonged storage of both fixed tissues and slide preparations, epitope redetection, sensitive immunoFISH-karyotyping. The described methodology provides unprecedented flexibility in laboratory work and significantly expands plant cyto-epigenetic research.