Antigen retrieval technique utilizing citrate buffer or urea solution for immunohistochemical demonstration (original) (raw)

Antigen retrieval technique utilizing citrate buffer or urea solution for immunohistochemical demonstration of androgen receptor in formalin-fixed paraffin sections.

Author(s): Shi SR, Chaiwun B, Young L, Cote RJ, Taylor CR

Publication: J Histochem Cytochem, 1993, Vol. 41, Page 1599

PubMed ID: 7691930PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to conduct a comparative assessment of nuclear androgen receptor (AR) immunostaining in formalin-fixed, paraffin embedded (FFPE) tissue fixed for 1 to 7 days and frozen tissue sections following microwave heating, enzyme digestion, or acid antigen retrieval approaches.

Conclusion of Paper

The authors report that optimal androgen receptor visualization with the lowest background was obtained in microwave-heated, dewaxed sections after pretreatment with pH 6.0, 5% urea or citrate buffer. This was qualitatively and quantitatively comparable to immunolocalization of androgen receptor in case-matched frozen biospecimens.

  1. Study Purpose

The purpose of this study was to conduct a comparative assessment of nuclear AR immunostaining in FFPE (fixed for 1-7 d) and frozen tissue sections following microwave heating, enzyme digestion, or acid antigen retrieval approaches. Tissues examined included prostate, breast, colon, lung, and stomach.

Summary of Findings:

The authors report that optimal androgen receptor visualization with the lowest background was obtained in microwave-heated, dewaxed sections after pretreatment with pH 6.0, 5% urea or citrate buffer. This was superior to pretreatment with saturated lead thiocyanate followed by microwave heating, and was qualitatively and quantitatively comparable to immunolocalization of androgen receptor in case-matched frozen biospecimens. Results held true for sections fixed for 24 hours or seven days. Enzymatic pretreatment with pepsin (5 min at 37 degrees C or 30 min at room temperature), trypsin (10 min at 37 degrees C), pronase E (5 min at 37 degrees C), trypsin and pronase E (sequentially for 5 min each at 37 degrees C) with no microwave heating resulted in less consistent staining and higher background.

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