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

S. R. Shi, B. Chaiwun, L. Young, Richard J Cote, C. R. Taylor

Research output: Contribution to journalArticle

243 Citations (Scopus)

Abstract

We developed a staining protocol for demonstration of androgen receptor (AR) in formalin-fixed, paraffin-embedded tissue sections. The method is based on the antigen retrieval microwave (MW) heating technique. Results are compared with different types of enzyme digestion pre-treatments. The strongest immunostaining signal and clearest background were obtained by MW heating of dewaxed paraffin sections in 5% urea or citrate buffer solution (pH 6); pure distilled water gave less consistent results. Enzymatic digestion with pepsin (0.05% in 2 N HCl) for 30 min at room temperature, or trypsin followed by pronase, or pronase digestion alone, also produced enhanced staining of AR in some cases, but there was more nonspecific background, and specific reactivity was less intense. The antigen retrieval MW method can be used to demonstrate AR epitope in prostate tissue after fixation in formalin for as long as 7 days. AR immunolocalization was also compared in frozen and paraffin sections processed from the same specimen of prostate carcinoma tissue and was found to be qualitatively and quantitatively similar. This study also provided new information concerning the basic principles of the antigen retrieval MW method that may be helpful in further development of this technique.

Original languageEnglish
Pages (from-to)1599-1604
Number of pages6
JournalJournal of Histochemistry and Cytochemistry
Volume41
Issue number11
StatePublished - Jan 1 1993
Externally publishedYes

Fingerprint

Androgen Receptors
Microwaves
Citric Acid
Paraffin
Formaldehyde
Urea
Buffers
Antigens
Digestion
Pronase
Heating
Prostate
Tissue Fixation
Staining and Labeling
Pepsin A
Frozen Sections
Trypsin
Epitopes
Carcinoma
Temperature

Keywords

  • Androgen receptor
  • Antigen retrieval technique
  • Enzyme digestion
  • Immunohistochemistry

ASJC Scopus subject areas

  • Cell Biology
  • Anatomy

Cite this

Antigen retrieval technique utilizing citrate buffer or urea solution for immunohistochemical demonstration of androgen receptor in formalin-fixed paraffin sections. / Shi, S. R.; Chaiwun, B.; Young, L.; Cote, Richard J; Taylor, C. R.

In: Journal of Histochemistry and Cytochemistry, Vol. 41, No. 11, 01.01.1993, p. 1599-1604.

Research output: Contribution to journalArticle

@article{8118ad3054a44c749ebe269e6d181305,
title = "Antigen retrieval technique utilizing citrate buffer or urea solution for immunohistochemical demonstration of androgen receptor in formalin-fixed paraffin sections",
abstract = "We developed a staining protocol for demonstration of androgen receptor (AR) in formalin-fixed, paraffin-embedded tissue sections. The method is based on the antigen retrieval microwave (MW) heating technique. Results are compared with different types of enzyme digestion pre-treatments. The strongest immunostaining signal and clearest background were obtained by MW heating of dewaxed paraffin sections in 5{\%} urea or citrate buffer solution (pH 6); pure distilled water gave less consistent results. Enzymatic digestion with pepsin (0.05{\%} in 2 N HCl) for 30 min at room temperature, or trypsin followed by pronase, or pronase digestion alone, also produced enhanced staining of AR in some cases, but there was more nonspecific background, and specific reactivity was less intense. The antigen retrieval MW method can be used to demonstrate AR epitope in prostate tissue after fixation in formalin for as long as 7 days. AR immunolocalization was also compared in frozen and paraffin sections processed from the same specimen of prostate carcinoma tissue and was found to be qualitatively and quantitatively similar. This study also provided new information concerning the basic principles of the antigen retrieval MW method that may be helpful in further development of this technique.",
keywords = "Androgen receptor, Antigen retrieval technique, Enzyme digestion, Immunohistochemistry",
author = "Shi, {S. R.} and B. Chaiwun and L. Young and Cote, {Richard J} and Taylor, {C. R.}",
year = "1993",
month = "1",
day = "1",
language = "English",
volume = "41",
pages = "1599--1604",
journal = "Journal of Histochemistry and Cytochemistry",
issn = "0022-1554",
publisher = "Histochemical Society Inc.",
number = "11",

}

TY - JOUR

T1 - Antigen retrieval technique utilizing citrate buffer or urea solution for immunohistochemical demonstration of androgen receptor in formalin-fixed paraffin sections

AU - Shi, S. R.

AU - Chaiwun, B.

AU - Young, L.

AU - Cote, Richard J

AU - Taylor, C. R.

PY - 1993/1/1

Y1 - 1993/1/1

N2 - We developed a staining protocol for demonstration of androgen receptor (AR) in formalin-fixed, paraffin-embedded tissue sections. The method is based on the antigen retrieval microwave (MW) heating technique. Results are compared with different types of enzyme digestion pre-treatments. The strongest immunostaining signal and clearest background were obtained by MW heating of dewaxed paraffin sections in 5% urea or citrate buffer solution (pH 6); pure distilled water gave less consistent results. Enzymatic digestion with pepsin (0.05% in 2 N HCl) for 30 min at room temperature, or trypsin followed by pronase, or pronase digestion alone, also produced enhanced staining of AR in some cases, but there was more nonspecific background, and specific reactivity was less intense. The antigen retrieval MW method can be used to demonstrate AR epitope in prostate tissue after fixation in formalin for as long as 7 days. AR immunolocalization was also compared in frozen and paraffin sections processed from the same specimen of prostate carcinoma tissue and was found to be qualitatively and quantitatively similar. This study also provided new information concerning the basic principles of the antigen retrieval MW method that may be helpful in further development of this technique.

AB - We developed a staining protocol for demonstration of androgen receptor (AR) in formalin-fixed, paraffin-embedded tissue sections. The method is based on the antigen retrieval microwave (MW) heating technique. Results are compared with different types of enzyme digestion pre-treatments. The strongest immunostaining signal and clearest background were obtained by MW heating of dewaxed paraffin sections in 5% urea or citrate buffer solution (pH 6); pure distilled water gave less consistent results. Enzymatic digestion with pepsin (0.05% in 2 N HCl) for 30 min at room temperature, or trypsin followed by pronase, or pronase digestion alone, also produced enhanced staining of AR in some cases, but there was more nonspecific background, and specific reactivity was less intense. The antigen retrieval MW method can be used to demonstrate AR epitope in prostate tissue after fixation in formalin for as long as 7 days. AR immunolocalization was also compared in frozen and paraffin sections processed from the same specimen of prostate carcinoma tissue and was found to be qualitatively and quantitatively similar. This study also provided new information concerning the basic principles of the antigen retrieval MW method that may be helpful in further development of this technique.

KW - Androgen receptor

KW - Antigen retrieval technique

KW - Enzyme digestion

KW - Immunohistochemistry

UR - http://www.scopus.com/inward/record.url?scp=0027484586&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027484586&partnerID=8YFLogxK

M3 - Article

C2 - 7691930

AN - SCOPUS:0027484586

VL - 41

SP - 1599

EP - 1604

JO - Journal of Histochemistry and Cytochemistry

JF - Journal of Histochemistry and Cytochemistry

SN - 0022-1554

IS - 11

ER -