Measurement of S phase duration in human epidermis using cyclin immunostaining and 3H-thymidine pulse labelling.
Cyclin/PCNA (proliferating cell nuclear antigen) is a nuclear protein strongly associated with DNA replication sites. Under methanol fixation, cyclin immunostaining might serve as a specific marker for S-phase cells. Associated with single labelling with 3H-thymidine, this immunodetection represents an easy method for measuring S-phase duration. Its application to six samples of normal human epidermis allowed us to measure an S phase of 9.7 +/- 0.3 h, a result in fair agreement with a previous estimate of 10.2 h obtained from a classical double-labelling technique with two different concentrations of 3H-thymidine.
Berlingin
E
E
Department of Dermatology, Hospital Erasme, Free University of Brussels, Belgium.
Heenen
M
M
Galand
P
P
D046508:Culture Techniques; D016213:Cyclins / Q000032:analysis* / Q000276:immunology; D000078404:Epidermal Cells; D006801:Humans; D016196:S Phase; D013936:Thymidine / Q000378:metabolism; D013997:Time Factors; D014316:Tritium
D016213:Cyclins; D014316:Tritium; D013936:Thymidine