Fluorophotometric quantitation of DNA in articular cartilage utilizing Hoechst 33258.
A sensitive fluorophotometric assay was developed for the measurement of DNA in articular cartilage. The tissue was digested with Proteinase K and dodecyl sodium sulfate, followed by analysis with Hoechst 33258 dye. DNA content was determined on both fresh and lyophilized material containing as little as 50 ng DNA. The results are comparable to values for other fluorophotometric and spectrophotometric methods reported in the literature. In addition, this method can be incorporated into existing methodology, allowing quantitation of specific glycosaminoglycans in the same cartilage sample in terms of DNA.
Lipman
J M
JM
Department of Pathology, State University of New York, Stony Brook 11794.
D000818:Animals; D006690:Bisbenzimidazole; D002358:Cartilage, Articular / Q000032:analysis*; D004247:DNA / Q000032:analysis*; D019286:Endopeptidase K; D011817:Rabbits; D012697:Serine Endopeptidases; D012967:Sodium Dodecyl Sulfate; D013050:Spectrometry, Fluorescence / Q000379:methods
D012967:Sodium Dodecyl Sulfate; D004247:DNA; D012697:Serine Endopeptidases; D019286:Endopeptidase K; D006690:Bisbenzimidazole