Improvement of immunodetection of the transcription factor C/EBP homologous protein by western blot

Accumulation of misfolded proteins within the endoplasmic reticulum (ER) induces an unfolded protein response (UPR) that either restores homeostasis or triggers apoptosis in case of adaptation failure. The three activated branches of UPR lead to IRE1-, PERK- and ATF6- dependent transcriptional induction of the gene encoding the transcription factor C/EBP homologous protein (CHOP) which plays an important role in apoptosis induction. In conventional immunoblotting conditions, detection of CHOP is a difficult task. Using a fixation step, we have optimized the detection of CHOP and this method provides a valuable tool to decipher CHOP involvement in UPR. [Display omitted] •Low retention of CHOP protein on nitrocellulose membrane during western blot.•Glutaraldehyde (GA) fixation increase reactivity to CHOP in western blot.•GA fixation decrease background.•GA fixation decrease non specific bands.