Strategies for improving the immunohistochemical staining of various intranuclear prognostic markers in formalin-paraffin sections: Androgen receptor, estrogen receptor, progesterone receptor, p53 protein, proliferating cell nuclear antigen, and Ki-67 antigen revealed by antigen retrieval techniques

Clive R. Taylor, Shan Rong Shi, Benjaporn Chaiwun, Lillian Young, S. Ashraf Imam, Richard J Cote

Research output: Contribution to journalArticle

253 Citations (Scopus)

Abstract

Different variations of the antigen retrieval technique using different retrieval solutions have been evaluated for their effectiveness in restoring the antigenicity of six intranuclear antigens, each of which is a potentially valuable prognostic indicator in formalin-fixed, paraffinembedded tissue sections. The results of immunohistochemical staining for estrogen receptor, progesterone receptor, androgen receptor, p53 protein, proliferating cell nuclear antigen, and Ki-67 antigen were compared following the different antigen retrieval approaches. The strongest immunostaining signal with the clearest background was obtained by microwave heating of dewaxed paraffin sections for 10 minutes in 0.05 mol/L glycine HCl (pH 3.5) or in citrate buffer solution (pH 6). Urea solution, distilled water, and lead thiocyanate solution yielded improvements with some antigens, but less consistently and less impressively than glycine HCl buffer or citrate buffer. Following antigen retrieval nuclear staining was sharply defined and could be achieved consistently in a variety of tissues after formalin fixation for as long as 7 days. The duration of fixation, however, was an important variable; generally, the longer the fixation time the more vigorous the retrieval procedure required. This study demonstrates the ability to stain a variety of intranuclear antigens, which are not readily demonstrable otherwise, in formalin-paraffin sections with a high degree of consistency and reproducibility. The availability of methods that are effective in paraffin sections may facilitate studies of the possible value of these markers as prognostic indicators for predicting the response of major tumors to different forms of therapy. This study also provided insight into the basic principles of the antigen retrieval method, which may be helpful in attempts to develop a more uniformly standardized technique applicable to many different antigen systems.

Original languageEnglish
Pages (from-to)263-270
Number of pages8
JournalHuman Pathology
Volume25
Issue number3
DOIs
StatePublished - Jan 1 1994
Externally publishedYes

Fingerprint

Ki-67 Antigen
Proliferating Cell Nuclear Antigen
Androgen Receptors
Progesterone Receptors
Estrogen Receptors
Paraffin
Formaldehyde
Staining and Labeling
Antigens
Buffers
Proteins
Citric Acid
Glycine
Antigenic Variation
Nuclear Antigens
Microwaves
Heating
Urea
Coloring Agents
Water

Keywords

  • androgen receptor
  • antigen retrieval
  • estrogen receptor
  • immunohistochemistry
  • Ki-67
  • p53 protein
  • progesterone receptor
  • proliferating cell nuclear antigen

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

@article{1e1a413ba661440e9b1a0e0e4777354f,
title = "Strategies for improving the immunohistochemical staining of various intranuclear prognostic markers in formalin-paraffin sections: Androgen receptor, estrogen receptor, progesterone receptor, p53 protein, proliferating cell nuclear antigen, and Ki-67 antigen revealed by antigen retrieval techniques",
abstract = "Different variations of the antigen retrieval technique using different retrieval solutions have been evaluated for their effectiveness in restoring the antigenicity of six intranuclear antigens, each of which is a potentially valuable prognostic indicator in formalin-fixed, paraffinembedded tissue sections. The results of immunohistochemical staining for estrogen receptor, progesterone receptor, androgen receptor, p53 protein, proliferating cell nuclear antigen, and Ki-67 antigen were compared following the different antigen retrieval approaches. The strongest immunostaining signal with the clearest background was obtained by microwave heating of dewaxed paraffin sections for 10 minutes in 0.05 mol/L glycine HCl (pH 3.5) or in citrate buffer solution (pH 6). Urea solution, distilled water, and lead thiocyanate solution yielded improvements with some antigens, but less consistently and less impressively than glycine HCl buffer or citrate buffer. Following antigen retrieval nuclear staining was sharply defined and could be achieved consistently in a variety of tissues after formalin fixation for as long as 7 days. The duration of fixation, however, was an important variable; generally, the longer the fixation time the more vigorous the retrieval procedure required. This study demonstrates the ability to stain a variety of intranuclear antigens, which are not readily demonstrable otherwise, in formalin-paraffin sections with a high degree of consistency and reproducibility. The availability of methods that are effective in paraffin sections may facilitate studies of the possible value of these markers as prognostic indicators for predicting the response of major tumors to different forms of therapy. This study also provided insight into the basic principles of the antigen retrieval method, which may be helpful in attempts to develop a more uniformly standardized technique applicable to many different antigen systems.",
keywords = "androgen receptor, antigen retrieval, estrogen receptor, immunohistochemistry, Ki-67, p53 protein, progesterone receptor, proliferating cell nuclear antigen",
author = "Taylor, {Clive R.} and Shi, {Shan Rong} and Benjaporn Chaiwun and Lillian Young and Imam, {S. Ashraf} and Cote, {Richard J}",
year = "1994",
month = "1",
day = "1",
doi = "10.1016/0046-8177(94)90198-8",
language = "English",
volume = "25",
pages = "263--270",
journal = "Human Pathology",
issn = "0046-8177",
publisher = "W.B. Saunders Ltd",
number = "3",

}

TY - JOUR

T1 - Strategies for improving the immunohistochemical staining of various intranuclear prognostic markers in formalin-paraffin sections

T2 - Androgen receptor, estrogen receptor, progesterone receptor, p53 protein, proliferating cell nuclear antigen, and Ki-67 antigen revealed by antigen retrieval techniques

AU - Taylor, Clive R.

AU - Shi, Shan Rong

AU - Chaiwun, Benjaporn

AU - Young, Lillian

AU - Imam, S. Ashraf

AU - Cote, Richard J

PY - 1994/1/1

Y1 - 1994/1/1

N2 - Different variations of the antigen retrieval technique using different retrieval solutions have been evaluated for their effectiveness in restoring the antigenicity of six intranuclear antigens, each of which is a potentially valuable prognostic indicator in formalin-fixed, paraffinembedded tissue sections. The results of immunohistochemical staining for estrogen receptor, progesterone receptor, androgen receptor, p53 protein, proliferating cell nuclear antigen, and Ki-67 antigen were compared following the different antigen retrieval approaches. The strongest immunostaining signal with the clearest background was obtained by microwave heating of dewaxed paraffin sections for 10 minutes in 0.05 mol/L glycine HCl (pH 3.5) or in citrate buffer solution (pH 6). Urea solution, distilled water, and lead thiocyanate solution yielded improvements with some antigens, but less consistently and less impressively than glycine HCl buffer or citrate buffer. Following antigen retrieval nuclear staining was sharply defined and could be achieved consistently in a variety of tissues after formalin fixation for as long as 7 days. The duration of fixation, however, was an important variable; generally, the longer the fixation time the more vigorous the retrieval procedure required. This study demonstrates the ability to stain a variety of intranuclear antigens, which are not readily demonstrable otherwise, in formalin-paraffin sections with a high degree of consistency and reproducibility. The availability of methods that are effective in paraffin sections may facilitate studies of the possible value of these markers as prognostic indicators for predicting the response of major tumors to different forms of therapy. This study also provided insight into the basic principles of the antigen retrieval method, which may be helpful in attempts to develop a more uniformly standardized technique applicable to many different antigen systems.

AB - Different variations of the antigen retrieval technique using different retrieval solutions have been evaluated for their effectiveness in restoring the antigenicity of six intranuclear antigens, each of which is a potentially valuable prognostic indicator in formalin-fixed, paraffinembedded tissue sections. The results of immunohistochemical staining for estrogen receptor, progesterone receptor, androgen receptor, p53 protein, proliferating cell nuclear antigen, and Ki-67 antigen were compared following the different antigen retrieval approaches. The strongest immunostaining signal with the clearest background was obtained by microwave heating of dewaxed paraffin sections for 10 minutes in 0.05 mol/L glycine HCl (pH 3.5) or in citrate buffer solution (pH 6). Urea solution, distilled water, and lead thiocyanate solution yielded improvements with some antigens, but less consistently and less impressively than glycine HCl buffer or citrate buffer. Following antigen retrieval nuclear staining was sharply defined and could be achieved consistently in a variety of tissues after formalin fixation for as long as 7 days. The duration of fixation, however, was an important variable; generally, the longer the fixation time the more vigorous the retrieval procedure required. This study demonstrates the ability to stain a variety of intranuclear antigens, which are not readily demonstrable otherwise, in formalin-paraffin sections with a high degree of consistency and reproducibility. The availability of methods that are effective in paraffin sections may facilitate studies of the possible value of these markers as prognostic indicators for predicting the response of major tumors to different forms of therapy. This study also provided insight into the basic principles of the antigen retrieval method, which may be helpful in attempts to develop a more uniformly standardized technique applicable to many different antigen systems.

KW - androgen receptor

KW - antigen retrieval

KW - estrogen receptor

KW - immunohistochemistry

KW - Ki-67

KW - p53 protein

KW - progesterone receptor

KW - proliferating cell nuclear antigen

UR - http://www.scopus.com/inward/record.url?scp=0028220224&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028220224&partnerID=8YFLogxK

U2 - 10.1016/0046-8177(94)90198-8

DO - 10.1016/0046-8177(94)90198-8

M3 - Article

C2 - 7512074

AN - SCOPUS:0028220224

VL - 25

SP - 263

EP - 270

JO - Human Pathology

JF - Human Pathology

SN - 0046-8177

IS - 3

ER -