A monoclonal antibody to rat asialoglycoprotein receptor that recognizes an epitope specific to its major subunit

Morizou Shimada, Motowo Mizuno, Tokurou Uesu, Junichirou Nasu, Hiroyuki Okada, Hiroyuki Shimomura, Kazuhide Yamamoto, Takao Tsuji, Yasushi Shiratori

Research output: Contribution to journalArticle

Abstract

We previously developed three mouse monoclonal antibodies (MoAbs) identifying the domain-specific plasma membrane proteins of rat hepatocytes. Based on the results of immunohistochemical and immunoblot analysis, one of them, clone 8D7, likely recognizes the rat asialoglycoprotein receptor (ASGPR), which consists of three polypeptide subunits: rat hepatic lectin (RHL)-1, RHL-2 and RHL-3. In this study, we examined the reactivity of MoAb 8D7 with these subunits in order to identify specifically the protein it recognizes. RHL-1 and RHL-2/3 were expressed in COS-1 mammalian kidney cells transfected with cloned cDNAs. Reactivity of MoAb 8D7 with these proteins was examined by flow cytometry, immunoblotting, and an immunofluorescence staining. By flow cytometry, MoAb 8D7 reacted with cell surface proteins on the COS-1 cells transfected with RHL-1 cDNA but not with RHL-2/3 cDNA. By immunoblotting, the antibody recognized proteins (molecular weights 45 and 96 kDa) in RHL-1-expressing cells but not in the RHL-2/3 cDNA-transfected cells. Immunoreactive staining with MoAb 8D7 was observed on the cell surface and in the cytoplasm of RHL-1-transfected COS-1 cells but not in cells expressing RHL-2/3. These findings indicate that MoAb 8D7 recognizes a subunit-specific epitope on RHL-1 of rat ASGPR.

Original languageEnglish
Pages (from-to)55-60
Number of pages6
JournalHepatology Research
Volume26
Issue number1
DOIs
Publication statusPublished - May 1 2003

Fingerprint

Asialoglycoprotein Receptor
Epitopes
Monoclonal Antibodies
Complementary DNA
COS Cells
Immunoblotting
Flow Cytometry
Membrane Proteins
Staining and Labeling
Proteins

Keywords

  • Asialoglycoprotein receptor
  • Epitope
  • Monoclonal antibody
  • RHL-18
  • Subunit

ASJC Scopus subject areas

  • Gastroenterology

Cite this

A monoclonal antibody to rat asialoglycoprotein receptor that recognizes an epitope specific to its major subunit. / Shimada, Morizou; Mizuno, Motowo; Uesu, Tokurou; Nasu, Junichirou; Okada, Hiroyuki; Shimomura, Hiroyuki; Yamamoto, Kazuhide; Tsuji, Takao; Shiratori, Yasushi.

In: Hepatology Research, Vol. 26, No. 1, 01.05.2003, p. 55-60.

Research output: Contribution to journalArticle

Shimada, M, Mizuno, M, Uesu, T, Nasu, J, Okada, H, Shimomura, H, Yamamoto, K, Tsuji, T & Shiratori, Y 2003, 'A monoclonal antibody to rat asialoglycoprotein receptor that recognizes an epitope specific to its major subunit', Hepatology Research, vol. 26, no. 1, pp. 55-60. https://doi.org/10.1016/S1386-6346(02)00329-7
Shimada, Morizou ; Mizuno, Motowo ; Uesu, Tokurou ; Nasu, Junichirou ; Okada, Hiroyuki ; Shimomura, Hiroyuki ; Yamamoto, Kazuhide ; Tsuji, Takao ; Shiratori, Yasushi. / A monoclonal antibody to rat asialoglycoprotein receptor that recognizes an epitope specific to its major subunit. In: Hepatology Research. 2003 ; Vol. 26, No. 1. pp. 55-60.
@article{d981aaaa2f9e4a298cebd9d1eeda5cf9,
title = "A monoclonal antibody to rat asialoglycoprotein receptor that recognizes an epitope specific to its major subunit",
abstract = "We previously developed three mouse monoclonal antibodies (MoAbs) identifying the domain-specific plasma membrane proteins of rat hepatocytes. Based on the results of immunohistochemical and immunoblot analysis, one of them, clone 8D7, likely recognizes the rat asialoglycoprotein receptor (ASGPR), which consists of three polypeptide subunits: rat hepatic lectin (RHL)-1, RHL-2 and RHL-3. In this study, we examined the reactivity of MoAb 8D7 with these subunits in order to identify specifically the protein it recognizes. RHL-1 and RHL-2/3 were expressed in COS-1 mammalian kidney cells transfected with cloned cDNAs. Reactivity of MoAb 8D7 with these proteins was examined by flow cytometry, immunoblotting, and an immunofluorescence staining. By flow cytometry, MoAb 8D7 reacted with cell surface proteins on the COS-1 cells transfected with RHL-1 cDNA but not with RHL-2/3 cDNA. By immunoblotting, the antibody recognized proteins (molecular weights 45 and 96 kDa) in RHL-1-expressing cells but not in the RHL-2/3 cDNA-transfected cells. Immunoreactive staining with MoAb 8D7 was observed on the cell surface and in the cytoplasm of RHL-1-transfected COS-1 cells but not in cells expressing RHL-2/3. These findings indicate that MoAb 8D7 recognizes a subunit-specific epitope on RHL-1 of rat ASGPR.",
keywords = "Asialoglycoprotein receptor, Epitope, Monoclonal antibody, RHL-18, Subunit",
author = "Morizou Shimada and Motowo Mizuno and Tokurou Uesu and Junichirou Nasu and Hiroyuki Okada and Hiroyuki Shimomura and Kazuhide Yamamoto and Takao Tsuji and Yasushi Shiratori",
year = "2003",
month = "5",
day = "1",
doi = "10.1016/S1386-6346(02)00329-7",
language = "English",
volume = "26",
pages = "55--60",
journal = "Hepatology Research",
issn = "1386-6346",
publisher = "Wiley-Blackwell",
number = "1",

}

TY - JOUR

T1 - A monoclonal antibody to rat asialoglycoprotein receptor that recognizes an epitope specific to its major subunit

AU - Shimada, Morizou

AU - Mizuno, Motowo

AU - Uesu, Tokurou

AU - Nasu, Junichirou

AU - Okada, Hiroyuki

AU - Shimomura, Hiroyuki

AU - Yamamoto, Kazuhide

AU - Tsuji, Takao

AU - Shiratori, Yasushi

PY - 2003/5/1

Y1 - 2003/5/1

N2 - We previously developed three mouse monoclonal antibodies (MoAbs) identifying the domain-specific plasma membrane proteins of rat hepatocytes. Based on the results of immunohistochemical and immunoblot analysis, one of them, clone 8D7, likely recognizes the rat asialoglycoprotein receptor (ASGPR), which consists of three polypeptide subunits: rat hepatic lectin (RHL)-1, RHL-2 and RHL-3. In this study, we examined the reactivity of MoAb 8D7 with these subunits in order to identify specifically the protein it recognizes. RHL-1 and RHL-2/3 were expressed in COS-1 mammalian kidney cells transfected with cloned cDNAs. Reactivity of MoAb 8D7 with these proteins was examined by flow cytometry, immunoblotting, and an immunofluorescence staining. By flow cytometry, MoAb 8D7 reacted with cell surface proteins on the COS-1 cells transfected with RHL-1 cDNA but not with RHL-2/3 cDNA. By immunoblotting, the antibody recognized proteins (molecular weights 45 and 96 kDa) in RHL-1-expressing cells but not in the RHL-2/3 cDNA-transfected cells. Immunoreactive staining with MoAb 8D7 was observed on the cell surface and in the cytoplasm of RHL-1-transfected COS-1 cells but not in cells expressing RHL-2/3. These findings indicate that MoAb 8D7 recognizes a subunit-specific epitope on RHL-1 of rat ASGPR.

AB - We previously developed three mouse monoclonal antibodies (MoAbs) identifying the domain-specific plasma membrane proteins of rat hepatocytes. Based on the results of immunohistochemical and immunoblot analysis, one of them, clone 8D7, likely recognizes the rat asialoglycoprotein receptor (ASGPR), which consists of three polypeptide subunits: rat hepatic lectin (RHL)-1, RHL-2 and RHL-3. In this study, we examined the reactivity of MoAb 8D7 with these subunits in order to identify specifically the protein it recognizes. RHL-1 and RHL-2/3 were expressed in COS-1 mammalian kidney cells transfected with cloned cDNAs. Reactivity of MoAb 8D7 with these proteins was examined by flow cytometry, immunoblotting, and an immunofluorescence staining. By flow cytometry, MoAb 8D7 reacted with cell surface proteins on the COS-1 cells transfected with RHL-1 cDNA but not with RHL-2/3 cDNA. By immunoblotting, the antibody recognized proteins (molecular weights 45 and 96 kDa) in RHL-1-expressing cells but not in the RHL-2/3 cDNA-transfected cells. Immunoreactive staining with MoAb 8D7 was observed on the cell surface and in the cytoplasm of RHL-1-transfected COS-1 cells but not in cells expressing RHL-2/3. These findings indicate that MoAb 8D7 recognizes a subunit-specific epitope on RHL-1 of rat ASGPR.

KW - Asialoglycoprotein receptor

KW - Epitope

KW - Monoclonal antibody

KW - RHL-18

KW - Subunit

UR - http://www.scopus.com/inward/record.url?scp=0042092005&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0042092005&partnerID=8YFLogxK

U2 - 10.1016/S1386-6346(02)00329-7

DO - 10.1016/S1386-6346(02)00329-7

M3 - Article

VL - 26

SP - 55

EP - 60

JO - Hepatology Research

JF - Hepatology Research

SN - 1386-6346

IS - 1

ER -