Immunocytochemical demonstration of erythropoietin in hypoxic human hepatoma cultures

C. Herkens; M. Wolff; J. Fandrey; F. Schuler; W. Jelkmann

doi:10.1007/BF00270051

Immunocytochemical demonstration of erythropoietin in hypoxic human hepatoma cultures

C. Herkens, M. Wolff, J. Fandrey, F. Schuler, W. Jelkmann^*

^*Corresponding author for this work

Institute of Physiology

University of Bonn

9 Citations (Scopus)

Abstract

The possibility of demonstrating erythropoietin at the light microscopic level was examined in homogeneous cultures of the erythropoietin-producing human hepatoma cell lines HepG2 and Hep3B. Immunoperoxidase staining was applied in combination with several mono- and polyclonal antibodies. Sufficiently strong colour responses were obtained with all three polyclonal antibodies and with one of three monoclonal antibodies raised against recombinant human erythropoietin. The staining intensity was increased in hypoxic versus non-hypoxic hepatoma cultures. Intracellular erythropoietin immunoreactivity was confirmed by Western blot analysis of HepG2 extracts. The effect of oxygen supply on erythropoietin gene expression was confirmed by competitive polymerase chain reaction of erythropoietin mRNA and by radioimmunoassay of secreted erythropoietin.

Original language	English
Journal	Histochemistry
Volume	100
Issue number	4
Pages (from-to)	303-309
Number of pages	7
ISSN	0301-5564
DOIs	https://doi.org/10.1007/BF00270051
Publication status	Published - 10.1993

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Academic Focus: Center for Brain, Behavior and Metabolism (CBBM)

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title = "Immunocytochemical demonstration of erythropoietin in hypoxic human hepatoma cultures",

abstract = "The possibility of demonstrating erythropoietin at the light microscopic level was examined in homogeneous cultures of the erythropoietin-producing human hepatoma cell lines HepG2 and Hep3B. Immunoperoxidase staining was applied in combination with several mono- and polyclonal antibodies. Sufficiently strong colour responses were obtained with all three polyclonal antibodies and with one of three monoclonal antibodies raised against recombinant human erythropoietin. The staining intensity was increased in hypoxic versus non-hypoxic hepatoma cultures. Intracellular erythropoietin immunoreactivity was confirmed by Western blot analysis of HepG2 extracts. The effect of oxygen supply on erythropoietin gene expression was confirmed by competitive polymerase chain reaction of erythropoietin mRNA and by radioimmunoassay of secreted erythropoietin.",

author = "C. Herkens and M. Wolff and J. Fandrey and F. Schuler and W. Jelkmann",

year = "1993",

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T1 - Immunocytochemical demonstration of erythropoietin in hypoxic human hepatoma cultures

AU - Herkens, C.

AU - Wolff, M.

AU - Fandrey, J.

AU - Schuler, F.

AU - Jelkmann, W.

PY - 1993/10

Y1 - 1993/10

N2 - The possibility of demonstrating erythropoietin at the light microscopic level was examined in homogeneous cultures of the erythropoietin-producing human hepatoma cell lines HepG2 and Hep3B. Immunoperoxidase staining was applied in combination with several mono- and polyclonal antibodies. Sufficiently strong colour responses were obtained with all three polyclonal antibodies and with one of three monoclonal antibodies raised against recombinant human erythropoietin. The staining intensity was increased in hypoxic versus non-hypoxic hepatoma cultures. Intracellular erythropoietin immunoreactivity was confirmed by Western blot analysis of HepG2 extracts. The effect of oxygen supply on erythropoietin gene expression was confirmed by competitive polymerase chain reaction of erythropoietin mRNA and by radioimmunoassay of secreted erythropoietin.

AB - The possibility of demonstrating erythropoietin at the light microscopic level was examined in homogeneous cultures of the erythropoietin-producing human hepatoma cell lines HepG2 and Hep3B. Immunoperoxidase staining was applied in combination with several mono- and polyclonal antibodies. Sufficiently strong colour responses were obtained with all three polyclonal antibodies and with one of three monoclonal antibodies raised against recombinant human erythropoietin. The staining intensity was increased in hypoxic versus non-hypoxic hepatoma cultures. Intracellular erythropoietin immunoreactivity was confirmed by Western blot analysis of HepG2 extracts. The effect of oxygen supply on erythropoietin gene expression was confirmed by competitive polymerase chain reaction of erythropoietin mRNA and by radioimmunoassay of secreted erythropoietin.

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DO - 10.1007/BF00270051

M3 - Journal articles

C2 - 8276645

AN - SCOPUS:0027420438

SN - 0301-5564

VL - 100

SP - 303

EP - 309

JO - Histochemistry

JF - Histochemistry

IS - 4

ER -

Immunocytochemical demonstration of erythropoietin in hypoxic human hepatoma cultures

Abstract

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