Use of pH 9.5 Tris-HCl buffer containing 5% urea for antigen retrieval immunohistochemistry

Shan-Rong-Shi, Richard J Cote, L. Young, S. A. Imam, C. R. Taylor

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Successful antigen retrieval (AR) immunohistochemistry is dependent on the temperature, heating time, and pH value of the AR solutions. There is no single standardized AR solution, however, that is suitable for all antibodies 'routinely' used in surgical pathology for immunostaining archival tissue sections. We tested a variety of AR solutions varying in pH value, chemical composition, and molarity. Based upon preliminary results, we compared three AR solutions: 0.1 M Tris-HCl buffer, pH 9.5, containing 5% urea, 0.1 M Tris- HCl buffer pH 9.5 without urea, and citrate buffer, pH 6.0. Each AR solution was tested with a panel of 34 antibodies using microwave heating for antigen retrieval. The heating conditions were standardized at 10 min and an automated stainer was used to standardize the immunostaining method. The Tris-HC1 containing urea was superior to pH 6.0 citrate buffer for 22 antibodies. In 12 cases, Tris-HCl with urea was also superior to Tris-HCl alone. In 12 cases, the intensity was similar for all three retrieval solutions. The staining obtained with Tris-HCl with urea was equal to or better than with pH 6.0 citrate buffer in all cases. The Tris-HC1 with urea solution is satisfactory for AR of most antibodies employed in routine surgical pathology.

Original languageEnglish
Pages (from-to)190-195
Number of pages6
JournalBiotechnic and Histochemistry
Volume71
Issue number4
StatePublished - Oct 4 1996
Externally publishedYes

Fingerprint

Tromethamine
Urea
Immunohistochemistry
Antigens
Citric Acid
Heating
Surgical Pathology
Buffers
Antibodies
Microwaves
Staining and Labeling
Temperature

Keywords

  • antibody
  • antigen
  • antigen retrieval
  • immunohistochemistry
  • microwave
  • paraffin section

ASJC Scopus subject areas

  • Anatomy
  • Applied Microbiology and Biotechnology

Cite this

Use of pH 9.5 Tris-HCl buffer containing 5% urea for antigen retrieval immunohistochemistry. / Shan-Rong-Shi, ; Cote, Richard J; Young, L.; Imam, S. A.; Taylor, C. R.

In: Biotechnic and Histochemistry, Vol. 71, No. 4, 04.10.1996, p. 190-195.

Research output: Contribution to journalArticle

Shan-Rong-Shi, , Cote, RJ, Young, L, Imam, SA & Taylor, CR 1996, 'Use of pH 9.5 Tris-HCl buffer containing 5% urea for antigen retrieval immunohistochemistry', Biotechnic and Histochemistry, vol. 71, no. 4, pp. 190-195.
Shan-Rong-Shi, ; Cote, Richard J ; Young, L. ; Imam, S. A. ; Taylor, C. R. / Use of pH 9.5 Tris-HCl buffer containing 5% urea for antigen retrieval immunohistochemistry. In: Biotechnic and Histochemistry. 1996 ; Vol. 71, No. 4. pp. 190-195.
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