Cell proliferation index determination by immunohistochemical detection of hCDC47 protein

Research output: Contribution to journalArticle

Abstract

A member of the human minichromosome maintenance complex protein family, hCDC47 (alias MCM7) has been identified as a component of the regulatory mechanism in cell proliferation. The expression of this protein, as determined by immunohistochemistry, was investigated to determine its application as a proliferation marker. A mouse monoclonal antibody (Clone 47DC141, NeoMarkers, Fremont CA) raised against recombinant hCDC47 protein was tested against a wide range of tissues. Immunoreaction patterns were determined in normal and neoplastic, human tissues, including skin, tonsils and lymph nodes, primary, and metastatic brain tumors. The protein was detected in the nuclei of both, normal and neoplastic proliferating cells. Similarly, we also examined the distribution of hCDC47 in normal rat and mouse tissues, and rodent and human tumors grown in nude mice.The pattern of immunolocalization was identical to that seen in human tissue, with positive nuclear immunoreaction readily identified in proliferating cells. Western immunoblot was carried out on extracts from PANC cells (human pancreatic adenocarcinoma cell line) to confirm the specificity of the protein. To correlate Ki67 protein immunoexpression with hCDC47 antibody reactivity, semiquantitative comparisons were carried out on parallel tissue sections. There was excellent correlation in the distribution pattern of the 2 markers, although hCDC47 was more sensitive.Thus this marker may have important clinical and research applications because of its activity in formalin-fixed, paraffin-embedded, proliferating, normal, and neoplastic tissue. More significantly, its application to animal tissue makes it a reliable and easy to use, proliferation marker for experimental studies.

Original languageEnglish (US)
Pages (from-to)278-282
Number of pages5
JournalApplied Immunohistochemistry and Molecular Morphology
Volume18
Issue number3
DOIs
StatePublished - May 1 2010
Externally publishedYes

Fingerprint

Cell Proliferation
Proteins
Minichromosome Maintenance Proteins
Palatine Tonsil
Cell Extracts
Recombinant Proteins
Nude Mice
Brain Neoplasms
Paraffin
Formaldehyde
Rodentia
Adenocarcinoma
Clone Cells
Lymph Nodes
Western Blotting
Immunohistochemistry
Monoclonal Antibodies
Cell Line
Skin
Antibodies

Keywords

  • HCDC47
  • Immunohistochemistry
  • MCM7
  • Proliferation marker

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology
  • Medical Laboratory Technology

Cite this

Cell proliferation index determination by immunohistochemical detection of hCDC47 protein. / Rojiani, Mumtaz V; Siemann, Dietmar W.; Rojiani, Amyn Mohammed.

In: Applied Immunohistochemistry and Molecular Morphology, Vol. 18, No. 3, 01.05.2010, p. 278-282.

Research output: Contribution to journalArticle

@article{2c35cddbe22c41e49dc66965c7f81710,
title = "Cell proliferation index determination by immunohistochemical detection of hCDC47 protein",
abstract = "A member of the human minichromosome maintenance complex protein family, hCDC47 (alias MCM7) has been identified as a component of the regulatory mechanism in cell proliferation. The expression of this protein, as determined by immunohistochemistry, was investigated to determine its application as a proliferation marker. A mouse monoclonal antibody (Clone 47DC141, NeoMarkers, Fremont CA) raised against recombinant hCDC47 protein was tested against a wide range of tissues. Immunoreaction patterns were determined in normal and neoplastic, human tissues, including skin, tonsils and lymph nodes, primary, and metastatic brain tumors. The protein was detected in the nuclei of both, normal and neoplastic proliferating cells. Similarly, we also examined the distribution of hCDC47 in normal rat and mouse tissues, and rodent and human tumors grown in nude mice.The pattern of immunolocalization was identical to that seen in human tissue, with positive nuclear immunoreaction readily identified in proliferating cells. Western immunoblot was carried out on extracts from PANC cells (human pancreatic adenocarcinoma cell line) to confirm the specificity of the protein. To correlate Ki67 protein immunoexpression with hCDC47 antibody reactivity, semiquantitative comparisons were carried out on parallel tissue sections. There was excellent correlation in the distribution pattern of the 2 markers, although hCDC47 was more sensitive.Thus this marker may have important clinical and research applications because of its activity in formalin-fixed, paraffin-embedded, proliferating, normal, and neoplastic tissue. More significantly, its application to animal tissue makes it a reliable and easy to use, proliferation marker for experimental studies.",
keywords = "HCDC47, Immunohistochemistry, MCM7, Proliferation marker",
author = "Rojiani, {Mumtaz V} and Siemann, {Dietmar W.} and Rojiani, {Amyn Mohammed}",
year = "2010",
month = "5",
day = "1",
doi = "10.1097/PAI.0b013e3181c6c949",
language = "English (US)",
volume = "18",
pages = "278--282",
journal = "Applied Immunohistochemistry and Molecular Morphology",
issn = "1541-2016",
publisher = "Lippincott Williams and Wilkins",
number = "3",

}

TY - JOUR

T1 - Cell proliferation index determination by immunohistochemical detection of hCDC47 protein

AU - Rojiani, Mumtaz V

AU - Siemann, Dietmar W.

AU - Rojiani, Amyn Mohammed

PY - 2010/5/1

Y1 - 2010/5/1

N2 - A member of the human minichromosome maintenance complex protein family, hCDC47 (alias MCM7) has been identified as a component of the regulatory mechanism in cell proliferation. The expression of this protein, as determined by immunohistochemistry, was investigated to determine its application as a proliferation marker. A mouse monoclonal antibody (Clone 47DC141, NeoMarkers, Fremont CA) raised against recombinant hCDC47 protein was tested against a wide range of tissues. Immunoreaction patterns were determined in normal and neoplastic, human tissues, including skin, tonsils and lymph nodes, primary, and metastatic brain tumors. The protein was detected in the nuclei of both, normal and neoplastic proliferating cells. Similarly, we also examined the distribution of hCDC47 in normal rat and mouse tissues, and rodent and human tumors grown in nude mice.The pattern of immunolocalization was identical to that seen in human tissue, with positive nuclear immunoreaction readily identified in proliferating cells. Western immunoblot was carried out on extracts from PANC cells (human pancreatic adenocarcinoma cell line) to confirm the specificity of the protein. To correlate Ki67 protein immunoexpression with hCDC47 antibody reactivity, semiquantitative comparisons were carried out on parallel tissue sections. There was excellent correlation in the distribution pattern of the 2 markers, although hCDC47 was more sensitive.Thus this marker may have important clinical and research applications because of its activity in formalin-fixed, paraffin-embedded, proliferating, normal, and neoplastic tissue. More significantly, its application to animal tissue makes it a reliable and easy to use, proliferation marker for experimental studies.

AB - A member of the human minichromosome maintenance complex protein family, hCDC47 (alias MCM7) has been identified as a component of the regulatory mechanism in cell proliferation. The expression of this protein, as determined by immunohistochemistry, was investigated to determine its application as a proliferation marker. A mouse monoclonal antibody (Clone 47DC141, NeoMarkers, Fremont CA) raised against recombinant hCDC47 protein was tested against a wide range of tissues. Immunoreaction patterns were determined in normal and neoplastic, human tissues, including skin, tonsils and lymph nodes, primary, and metastatic brain tumors. The protein was detected in the nuclei of both, normal and neoplastic proliferating cells. Similarly, we also examined the distribution of hCDC47 in normal rat and mouse tissues, and rodent and human tumors grown in nude mice.The pattern of immunolocalization was identical to that seen in human tissue, with positive nuclear immunoreaction readily identified in proliferating cells. Western immunoblot was carried out on extracts from PANC cells (human pancreatic adenocarcinoma cell line) to confirm the specificity of the protein. To correlate Ki67 protein immunoexpression with hCDC47 antibody reactivity, semiquantitative comparisons were carried out on parallel tissue sections. There was excellent correlation in the distribution pattern of the 2 markers, although hCDC47 was more sensitive.Thus this marker may have important clinical and research applications because of its activity in formalin-fixed, paraffin-embedded, proliferating, normal, and neoplastic tissue. More significantly, its application to animal tissue makes it a reliable and easy to use, proliferation marker for experimental studies.

KW - HCDC47

KW - Immunohistochemistry

KW - MCM7

KW - Proliferation marker

UR - http://www.scopus.com/inward/record.url?scp=77951718574&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77951718574&partnerID=8YFLogxK

U2 - 10.1097/PAI.0b013e3181c6c949

DO - 10.1097/PAI.0b013e3181c6c949

M3 - Article

VL - 18

SP - 278

EP - 282

JO - Applied Immunohistochemistry and Molecular Morphology

JF - Applied Immunohistochemistry and Molecular Morphology

SN - 1541-2016

IS - 3

ER -