Specific demonstration of ribonucleic acid by chemical bromination and immunohistochemistry. | Zendy

G. Harms | Zendy; M. J. Hardonk | Zendy

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Specific demonstration of ribonucleic acid by chemical bromination and immunohistochemistry.

Author(s) -

G. Harms,

M. J. Hardonk

Publication year - 1989

Publication title -

journal of histochemistry and cytochemistry

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.971

H-Index - 124

eISSN - 1551-5044

pISSN - 0022-1554

DOI - 10.1177/37.4.2466888

Subject(s) - halogenation , bromine , chemistry , rna , staining , potassium bromide , sodium bisulfite , ribonuclease , rnase p , iodine , biochemistry , chromatography , organic chemistry , biology , genetics , gene

In this report we describe a specific staining procedure for detection of ribonucleic acid (RNA), based on bromination of uracil and subsequent immunohistochemical visualization of 5-bromouracil in RNA. This method is applicable for both cryostat and glycol methacrylate (GMA)-embedded sections. Cryostat sections must be fixed in formaldehyde, whereas tissue pieces to be embedded in GMA are fixed in cold acetone. Before bromination, sections must be treated with trypsin. Bromination was performed in a solution of bromine in potassium bromide. After bromination, excess bromine was removed with sodium bisulfite. The monoclonal antibody MoBu-1 specifically bound to brominated RNA. Ribonuclease digestion, in contrast to deoxyribonuclease digestion, abolished staining. This method makes possible precise localization of RNA, especially well demonstrated in plastic-embedded sections.

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