ISSN:
1573-0603
Keywords:
DNA synthesis
;
erythropoietin
;
cell proliferation
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Summary Methods are presented for the assay of [3H]thymidine incorporation into anchorage-dependent cells (3T3 fibroblasts) using 96-well microplates. The plated cells remain in the wells throughout the procedure. In the final step, trichloroacetic acid-precipitated materials are hydrolyzed directly in the wells and aliquots are taken for counting. The same method is used for anchorage-independent cells (K562 or liver erythroid cells) but the incubations are carried out in 96-well-matched micro test tubes. The cells are washed from nonincorporated thymidine by centrifugation, and DNA is hydrolyzed as indicated for the anchorage-dependent cells. These methods avoid the use of expensive and time-consuming cell harvesting machines and the problems of quantitative transfer and filtration of precipitates. At the same time, the small incubation volumes decrease the cost of reagents, growth factors, and tissue culture media.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF01404641
Permalink