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A flow cytometric assay for the determination of cell proliferation with a monoclonal antibody directed against DNA‐methyltransferase
Author(s) -
Neubauer Andreas,
Serke Stefan,
Siegert Wolfgang,
Kroll Werner,
Musch Reinhard,
Huhn Dieter
Publication year - 1989
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.1989.tb04311.x
Subject(s) - monoclonal antibody , flow cytometry , microbiology and biotechnology , dna , monoclonal , antibody , virology , biology , immunology , chemistry , genetics
Summary The enzyme DNA‐methyltransferase is responsible for the methylation of a newly synthesized DNA‐strand. A monoclonal antibody directed against DNA‐methyltransferase was used to determine cell proliferation by means of flow cytometry. The reactivity of DNA‐methyltransferase antibody was compared with the known proliferation markers transferrin‐receptor and Ki67. All three methods showed comparable reactivity with the erythroblastic cell line K562 (86%, 81%, 76% respectively). In a second set of experiments peripheral blood mononuclear cells were stimulated with phytohaemagglutinin; in three experiments, a mean of 63% of the cells reacted with DNA‐methyltransferase antibody after 72 h of culture as compared to a mean of 6% in the case of unstimulated control cells. HL60 cells were incubated with DMSO and harvested on day 5 of culture. The results obtained show that in differentiated cells the fraction positive with DNA‐methyltransferase antibody decreased to levels below 10%. It is concluded that the technique described is a fast and easy method for the flow cytometric determination of cellular proliferation.