Recognition of murine integrin β1 by a rat anti-stromal cell monoclonal antibody

X. Wu, K. Miyake, Kay L Medina, P. W. Kincade, J. M. Gimble

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Previous studies with the rat monoclonal antibody KMI6 had localized its antigen in vivo to a discrete subpopulation of marrow stromal cells. The KMI6 antigen has now been identified as the murine homolog of integrin β1 by amino acid sequence analysis and by cross-reactivity with antiserum to the avian integrin β1. The relative tissue abundance of murine integrin β1 was determined by Western blot. Although immunoperoxidase staining of fixed murine hematopoietic tissues demonstrated an abundance of intracellular β1, few primary-derived cells of lymphohematopoietic origin were surface positive as assessed by immunofluorescence and flow cytometry. Fetal erythroblasts provided the only exception. In contrast, the antigen was readily-detected on the surface of several cultured cell lines in association with a variety of α chains. The biochemical properties of the surface labeled murine integrin β1 were similar to those of its human counterpart, exhibiting an altered electrophoretic migration under reduced conditions or following N-glycanase treatment. The antibody recognition of the protein was insensitive to glycosylation state, presence of divalent cations, detergents, or transfer to a nitrocellulose membrane. However, on Western blot, the epitope was lost on reduction of the protein, suggesting that it is conformation dependent. These data indicate that although KMI6 epitope is widely distributed, its surface expression in vivo may be restricted within lymphohemopoietic tissues.

Original languageEnglish (US)
Pages (from-to)409-416
Number of pages8
JournalHybridoma
Volume13
Issue number5
StatePublished - 1994
Externally publishedYes

Fingerprint

Stromal Cells
Integrins
Monoclonal Antibodies
Antigens
Epitopes
Western Blotting
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Erythroblasts
Collodion
Surface Properties
Protein Sequence Analysis
Divalent Cations
Glycosylation
Detergents
Fluorescent Antibody Technique
Immune Sera
Cultured Cells
Flow Cytometry
Proteins
Bone Marrow

ASJC Scopus subject areas

  • Immunology
  • Genetics

Cite this

Wu, X., Miyake, K., Medina, K. L., Kincade, P. W., & Gimble, J. M. (1994). Recognition of murine integrin β1 by a rat anti-stromal cell monoclonal antibody. Hybridoma, 13(5), 409-416.

Recognition of murine integrin β1 by a rat anti-stromal cell monoclonal antibody. / Wu, X.; Miyake, K.; Medina, Kay L; Kincade, P. W.; Gimble, J. M.

In: Hybridoma, Vol. 13, No. 5, 1994, p. 409-416.

Research output: Contribution to journalArticle

Wu, X, Miyake, K, Medina, KL, Kincade, PW & Gimble, JM 1994, 'Recognition of murine integrin β1 by a rat anti-stromal cell monoclonal antibody', Hybridoma, vol. 13, no. 5, pp. 409-416.
Wu, X. ; Miyake, K. ; Medina, Kay L ; Kincade, P. W. ; Gimble, J. M. / Recognition of murine integrin β1 by a rat anti-stromal cell monoclonal antibody. In: Hybridoma. 1994 ; Vol. 13, No. 5. pp. 409-416.
@article{8479f9a82d1b47bfb214370522fa4d17,
title = "Recognition of murine integrin β1 by a rat anti-stromal cell monoclonal antibody",
abstract = "Previous studies with the rat monoclonal antibody KMI6 had localized its antigen in vivo to a discrete subpopulation of marrow stromal cells. The KMI6 antigen has now been identified as the murine homolog of integrin β1 by amino acid sequence analysis and by cross-reactivity with antiserum to the avian integrin β1. The relative tissue abundance of murine integrin β1 was determined by Western blot. Although immunoperoxidase staining of fixed murine hematopoietic tissues demonstrated an abundance of intracellular β1, few primary-derived cells of lymphohematopoietic origin were surface positive as assessed by immunofluorescence and flow cytometry. Fetal erythroblasts provided the only exception. In contrast, the antigen was readily-detected on the surface of several cultured cell lines in association with a variety of α chains. The biochemical properties of the surface labeled murine integrin β1 were similar to those of its human counterpart, exhibiting an altered electrophoretic migration under reduced conditions or following N-glycanase treatment. The antibody recognition of the protein was insensitive to glycosylation state, presence of divalent cations, detergents, or transfer to a nitrocellulose membrane. However, on Western blot, the epitope was lost on reduction of the protein, suggesting that it is conformation dependent. These data indicate that although KMI6 epitope is widely distributed, its surface expression in vivo may be restricted within lymphohemopoietic tissues.",
author = "X. Wu and K. Miyake and Medina, {Kay L} and Kincade, {P. W.} and Gimble, {J. M.}",
year = "1994",
language = "English (US)",
volume = "13",
pages = "409--416",
journal = "Monoclonal Antibodies in Immunodiagnosis and Immunotherapy",
issn = "2167-9436",
publisher = "Mary Ann Liebert Inc.",
number = "5",

}

TY - JOUR

T1 - Recognition of murine integrin β1 by a rat anti-stromal cell monoclonal antibody

AU - Wu, X.

AU - Miyake, K.

AU - Medina, Kay L

AU - Kincade, P. W.

AU - Gimble, J. M.

PY - 1994

Y1 - 1994

N2 - Previous studies with the rat monoclonal antibody KMI6 had localized its antigen in vivo to a discrete subpopulation of marrow stromal cells. The KMI6 antigen has now been identified as the murine homolog of integrin β1 by amino acid sequence analysis and by cross-reactivity with antiserum to the avian integrin β1. The relative tissue abundance of murine integrin β1 was determined by Western blot. Although immunoperoxidase staining of fixed murine hematopoietic tissues demonstrated an abundance of intracellular β1, few primary-derived cells of lymphohematopoietic origin were surface positive as assessed by immunofluorescence and flow cytometry. Fetal erythroblasts provided the only exception. In contrast, the antigen was readily-detected on the surface of several cultured cell lines in association with a variety of α chains. The biochemical properties of the surface labeled murine integrin β1 were similar to those of its human counterpart, exhibiting an altered electrophoretic migration under reduced conditions or following N-glycanase treatment. The antibody recognition of the protein was insensitive to glycosylation state, presence of divalent cations, detergents, or transfer to a nitrocellulose membrane. However, on Western blot, the epitope was lost on reduction of the protein, suggesting that it is conformation dependent. These data indicate that although KMI6 epitope is widely distributed, its surface expression in vivo may be restricted within lymphohemopoietic tissues.

AB - Previous studies with the rat monoclonal antibody KMI6 had localized its antigen in vivo to a discrete subpopulation of marrow stromal cells. The KMI6 antigen has now been identified as the murine homolog of integrin β1 by amino acid sequence analysis and by cross-reactivity with antiserum to the avian integrin β1. The relative tissue abundance of murine integrin β1 was determined by Western blot. Although immunoperoxidase staining of fixed murine hematopoietic tissues demonstrated an abundance of intracellular β1, few primary-derived cells of lymphohematopoietic origin were surface positive as assessed by immunofluorescence and flow cytometry. Fetal erythroblasts provided the only exception. In contrast, the antigen was readily-detected on the surface of several cultured cell lines in association with a variety of α chains. The biochemical properties of the surface labeled murine integrin β1 were similar to those of its human counterpart, exhibiting an altered electrophoretic migration under reduced conditions or following N-glycanase treatment. The antibody recognition of the protein was insensitive to glycosylation state, presence of divalent cations, detergents, or transfer to a nitrocellulose membrane. However, on Western blot, the epitope was lost on reduction of the protein, suggesting that it is conformation dependent. These data indicate that although KMI6 epitope is widely distributed, its surface expression in vivo may be restricted within lymphohemopoietic tissues.

UR - http://www.scopus.com/inward/record.url?scp=0028114178&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028114178&partnerID=8YFLogxK

M3 - Article

VL - 13

SP - 409

EP - 416

JO - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

JF - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

SN - 2167-9436

IS - 5

ER -