Dual staining of lymphocyte membrane antigens with colloidal gold and biotinylated horseradish peroxidase

C. E. Gibson; I. A. Penttila; A. S Y Leong; J. Milios; H. Zola; J. T. LaBrooy

doi:10.1016/0022-1759(94)90075-2

Dual staining of lymphocyte membrane antigens with colloidal gold and biotinylated horseradish peroxidase

C. E. Gibson, I. A. Penttila, A. S Y Leong, J. Milios, H. Zola, J. T. LaBrooy

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Abstract

Double immunoenzymatic labelling procedures for the localization of antigens on cells in tissue sections using horseradish peroxidase (HRP) and alkaline phosphatase have been described previously, but mainly for detecting antigens on different cells. With this type of staining when two antigens are present on the same cell, an optimal colour combination that shows a high contrast between the basic colour of each enzyme substrate product is difficult to achieve and the interpretation of their mixed colour intermediate is subjective. We present a method for the simultaneous demonstration of two antigens on the same cell. The method can be used to label either single cells in suspension, or cells in paraffin fixed, tissue using a combination of a particulate label, colloidal immunogold-silver, and an enzymatic label HRP-DAB. The method is easy to perform and utilises commercially available staining kits.

Original language	English
Pages (from-to)	65-73
Number of pages	9
Journal	Journal of Immunological Methods
Volume	167
Issue number	1-2
DOIs	https://doi.org/10.1016/0022-1759(94)90075-2
Publication status	Published - 3 Jan 1994

Keywords

Dual immunohistochemical labeling
Horseradish peroxidase
Immunogold-silver

ASJC Scopus subject areas

Immunology and Allergy
Immunology

Access to Document

10.1016/0022-1759(94)90075-2

Link to publication in Scopus

Cite this

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title = "Dual staining of lymphocyte membrane antigens with colloidal gold and biotinylated horseradish peroxidase",

abstract = "Double immunoenzymatic labelling procedures for the localization of antigens on cells in tissue sections using horseradish peroxidase (HRP) and alkaline phosphatase have been described previously, but mainly for detecting antigens on different cells. With this type of staining when two antigens are present on the same cell, an optimal colour combination that shows a high contrast between the basic colour of each enzyme substrate product is difficult to achieve and the interpretation of their mixed colour intermediate is subjective. We present a method for the simultaneous demonstration of two antigens on the same cell. The method can be used to label either single cells in suspension, or cells in paraffin fixed, tissue using a combination of a particulate label, colloidal immunogold-silver, and an enzymatic label HRP-DAB. The method is easy to perform and utilises commercially available staining kits.",

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T1 - Dual staining of lymphocyte membrane antigens with colloidal gold and biotinylated horseradish peroxidase

AU - Gibson, C. E.

AU - Penttila, I. A.

AU - Leong, A. S Y

AU - Milios, J.

AU - Zola, H.

AU - LaBrooy, J. T.

PY - 1994/1/3

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AB - Double immunoenzymatic labelling procedures for the localization of antigens on cells in tissue sections using horseradish peroxidase (HRP) and alkaline phosphatase have been described previously, but mainly for detecting antigens on different cells. With this type of staining when two antigens are present on the same cell, an optimal colour combination that shows a high contrast between the basic colour of each enzyme substrate product is difficult to achieve and the interpretation of their mixed colour intermediate is subjective. We present a method for the simultaneous demonstration of two antigens on the same cell. The method can be used to label either single cells in suspension, or cells in paraffin fixed, tissue using a combination of a particulate label, colloidal immunogold-silver, and an enzymatic label HRP-DAB. The method is easy to perform and utilises commercially available staining kits.

KW - Dual immunohistochemical labeling

KW - Horseradish peroxidase

KW - Immunogold-silver

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