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

ABSTRACT:

Background

Protein detection on large somatic chromosomes typically includes paraformaldehyde fixation and squashing of enzymatically softened root tips in a buffer. It often suffers from chromosome clumping, poor chromosome morphology, non-specific fluorescence, insufficient immunoreactivity, which collectively reduce the credibility of immunolabeling, hindering its effective combination with fluorescence in situ hybridization (FISH). Material harvesting and pre-detection steps must be completed within a short time, usually one day, which complicates research. The aim of this study was to develop a simple efficient squash-based protocol for technically demanding formaldehyde-fixed large chromosomes/nuclei (Allium, Scilla, Tradescantia), that ensures: long-term storage of the fixed

SUBMITTER: Golczyk H 

PROVIDER: S-EPMC12459044 | biostudies-literature | 2025 Sep

REPOSITORIES: biostudies-literature