Nonradioactive in situ Hybridization Techniques for Routinely Prepared Pathology Specimens and Cultured Cells
作者:
yunFang,
LukGordon D.,
GesellMark S.,
期刊:
Journal of Histotechnology
(Taylor Available online 1994)
卷期:
Volume 17,
issue 4
页码: 313-319
ISSN:0147-8885
年代: 1994
DOI:10.1179/his.1994.17.4.313
出版商: Taylor&Francis
关键词: beta-actin;c-myc;cell cultures;digoxigenin-labeling;in situ hybridization;ornithine decarboxylase;pathology archives;random primer extension
数据来源: Taylor
摘要:
AbstractWe report a modified method for non-radioactive in situ hybridization suitable for semiquantitative analysis of specific mRNA sequences in fixed cultured cells or formalin fixed, paraffin embedded tissue sections. Our modifications include random primer incorporation of digoxigenin-label into DNA probe; specimen treatment with proteinase K; heating of specimen and hybridization solution to 100°C for 10 min followed by hybridization at 42°C for 18 hr; and improved immunologic detection with an antibody against digoxigenin conjugate. Visual results were obtained within 3 days. Probe sizes as large as 1.8 kb resulted in good specific signals with minimal background. Specific histologic localization can be semiquantitated under a brightfield microscope linked to a computer aided densitometry system. (The J Histotechnol17:313, 1994)
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