Ce. Gibson et al., DUAL STAINING OF LYMPHOCYTE MEMBRANE-ANTIGENS WITH COLLOIDAL GOLD ANDBIOTINYLATED HORSERADISH-PEROXIDASE, Journal of immunological methods, 167(1-2), 1994, pp. 65-73
Double immunoenzymatic labelling procedures for the localization of an
tigens on cells in tissue sections using horseradish peroxidase (HRP)
and alkaline phosphatase have been described previously, but mainly fo
r detecting antigens on different cells. With this type of staining wh
en two antigens are present on the same cell, an optimal colour combin
ation that shows a high contrast between the basic colour of each enzy
me substrate product is difficult to achieve and the interpretation of
their mixed colour intermediate is subjective. We present a method fo
r the simultaneous demonstration of two antigens on the same cell. The
method can be used to label either single cells in suspension, or cel
ls in paraffin fixed tissue, using a combination of a particulate labe
l, colloidal immunogold-silver, and an enzymatic label HRP-DAB. The me
thod is easy to perform and utilises commerically available staining k
its.