| Autor: |
Beisker W, Dolbeare F, Gray JW |
| Jazyk: |
angličtina |
| Zdroj: |
Cytometry [Cytometry] 1987 Mar; Vol. 8 (2), pp. 235-9. |
| DOI: |
10.1002/cyto.990080218 |
| Abstrakt: |
This report describes an improved immunochemical procedure to stain cells in suspension for incorporated bromodeoxyuridine (BrdUrd) and total DNA content. The procedure consists of five steps: chromatin proteins are extracted by treating with 0.1 M HCl and 0.7% Triton X-100 to facilitate DNA denaturation and to minimize nonspecific staining; cellular DNA is denatured by heating to 100 degrees C in distilled water; BrdUrd in single-stranded DNA (ssDNA) is stained using an immunochemical procedure; autofluorescence is reduced using sodium borohydride (NaBH4); and DNA is stained with the fluorescent dye propidium iodide. With this procedure, the BrdUrd incorporated by CHO cells during periods as short as a few seconds can be detected using flow cytometry. In addition, the stoichiometry of the immunofluorescent staining procedure is high. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
|
Plný text