TY - JOUR
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.
N1 - Funding Information:
This project was funded by the Australian National Health and Medical Research Council and undertaken whilst Dr. I. Penttila was in receipt of a Royal Adelaide Hospital Florey post doctoral research fellowship. We are grateful to the volunteers who donated blood, and for the assistance of Peter Gillum and Loretta Flego in the tissue staining, flow cytometry analysis, and antibody biotinylation.
PY - 1994/1/3
Y1 - 1994/1/3
N2 - 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.
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
UR - http://www.scopus.com/inward/record.url?scp=0027953622&partnerID=8YFLogxK
U2 - 10.1016/0022-1759(94)90075-2
DO - 10.1016/0022-1759(94)90075-2
M3 - Article
C2 - 7508481
AN - SCOPUS:0027953622
VL - 167
SP - 65
EP - 73
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
SN - 0022-1759
IS - 1-2
ER -