A rapid and simple immunoperoxidase staining procedure for blood and bone marrow samples.
A fast and simple indirect immunoperoxidase staining technique is described, which can also be used for the characterization of cells with high endogenous peroxidase activity such as many haemopoietic cells. It is based on the combination of a newly developed glucose-oxidase plus glucose procedure for the inhibition of endogenous peroxidase activity with a standard 2 or 3 layer immunoperoxidase staining protocol. Glucose-oxidase plus glucose mixture completely inhibits endogenous peroxidase activity without having a detectable deleterious effect on any of the cellular antigens so far studied. In many instances this permits the use of only 1 layer of horseradish peroxidase (HRP)-labelled antibodies. The glucose-oxidase plus glucose mixture can also be added to cells together with the HRP-labelled antibody solution without losing its inhibitory effect for endogenous peroxidase activity and without leading to a visually detectable loss of the activity of the HRP conjugate. Consequently, the separate incubation step previously necessary for the inhibition of endogenous peroxidase activity becomes superfluous.
U
U
Köller
H
H
Stockinger
O
O
Majdic
P
P
Bettelheim
W
W
Knapp
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000032:analysis; D001773:Blood Cells / Q000201:enzymology / Q000276:immunology*; D001853:Bone Marrow / Q000201:enzymology / Q000276:immunology*; D005404:Fixatives; D006098:Granulocytes / Q000201:enzymology / Q000276:immunology; D006801:Humans; D007124:Immunoenzyme Techniques; D008214:Lymphocytes / Q000201:enzymology / Q000276:immunology; D008533:Megakaryocytes / Q000201:enzymology / Q000276:immunology; D009000:Monocytes / Q000201:enzymology / Q000276:immunology; D009504:Neutrophils / Q000201:enzymology / Q000276:immunology; D010544:Peroxidases / Q000378:metabolism*; D013194:Staining and Labeling
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D005404:Fixatives; D010544:Peroxidases