Fixation requirements for the immunohistochemical reactivity of PCNA antibody PC10 on cryostat sections.
In contrast to that in paraffin-embedded tissue, the reactivity of monoclonal PCNA antibody PC10 on cryostat sections requires a special fixation procedure as the target epitope is seemingly not accessible to its antibody. A panel of 18 fixation protocols was investigated. Chilled methanol or acetone, or PLP (paraformaldehyde-lysine-periodate) was found to be unsuitable for skin preparations. A two-step fixation protocol was developed for normal skin and basal cell carcinomas. They were fixed first in 3.4% buffered formaldehyde, followed by fixation in 2:1 v/v ethanol-acetic acid. Following this fixation regime, cryostat sections displayed the same PCNA/PC10 labelling pattern as paraffin sections of formalin-fixed tissue.
Baum
H P
HP
Department of Dermatology, University of Saarland, Homburg/Saar, Germany.
Reichrath
J
J
Theobald
A
A
Schock
G
G
D000911:Antibodies, Monoclonal; D002280:Basal Cell Carcinoma / Q000737:chemistry* / Q000473:pathology; D016616:Cryoultramicrotomy; D005629:Frozen Sections; D006801:Humans; D007124:Immunoenzyme Techniques; D016612:Paraffin Embedding; D018809:Proliferating Cell Nuclear Antigen / Q000032:analysis* / Q000276:immunology; D012867:Skin / Q000737:chemistry / Q000473:pathology; D012878:Skin Neoplasms / Q000737:chemistry* / Q000473:pathology; D016707:Tissue Fixation / Q000379:methods*
D000911:Antibodies, Monoclonal; D018809:Proliferating Cell Nuclear Antigen