Abstract
Antigen modification and denaturation are recognized causes of false negatives in immunostaining. Specimens that have been stored for an extended period at room temperature show decreased immunoreactivity and may mislead the diagnosis. Studies of the molecular targeting of drugs often involve immunostaining of previous samples and, in some situations, only unstained specimens can be used. The present study aimed to develop an effective staining method to recover antigen activation in unstained specimens stored for an extended period by using ethylene-diamine-tetraacetic acid (EDTA) buffer solution with boric acid. We compared several commonly used antigen retrieval solutions and found that Tris-borate-EDTA (TBE) buffer solution with a pH ≥8.3 provided sufficient antigen retrieval. However, pH values higher than 8.3 (9.0, 10.0, and 11.0) frequently caused severe tissue damage. Thus, TBE with pH 8.3 was the most suitable antigen retrieval solution for recovering the antigenicity of specimens stored for an extended period. This procedure may allow useful immunohistochemical information, even from sections that have been stored for an extended period.
Highlights
Antigen modification and denaturation are recognized causes of false-negative reactions in immunostaining
The present study aimed to develop an effective staining method that improves antigen activation of old unstained specimens by using an ethylene-diamine-tetraacetic acid (EDTA) buffer solution with boric acid
We examined TBE with a pH of 9.0, 10, and 11
Summary
Antigen modification and denaturation are recognized causes of false-negative reactions in immunostaining. The effectiveness of a thermal treatment that uses boric acid buffer solution for DNA analysis with polymerase chain reaction (PCR) was reported, and the efficient separation performance of an ethylene-diamine-tetraacetic acid (EDTA) buffer solution containing Tris-saltbased borax has garnered attention as an electrophoresis buffer solution for DNA and RNA [8,9]. We hypothesized that this buffer might be effective in immunostaining. The present study aimed to develop an effective staining method that improves antigen activation of old unstained specimens by using an EDTA buffer solution with boric acid
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