Tyramine amplification technique in routine immunohistochemistry.
Signal amplification in immunohistochemistry via binding of biotinylated tyramine to proteins near the site of peroxidase-labeled antibodies is a promising new technique, but studies investigating a wide range of markers are lacking. The tyramine amplification technique (TAT) was investigated on 85 antibodies using a simple and fast protocol, and TAT results were compared to those obtained with conventional immunohistochemistry. Using TAT, most of the markers could be 5- to 50-fold further diluted and still showed identical staining results compared with standard stainings (maximal 500-fold). However, the variable reactivity of the different markers with TAT underlines the need for individual testing of every antibody to determine the optimal dilution. Some antibodies against cell adhesion molecules could be demonstrated for the first time in archival, formalin-fixed tissue sections. TAT, if carefully evaluated, offers a revolutionary improvement for modern immunostaining, either to increase sensitivity or primary antibody dilutions (cost reduction). From a methodological point of view, immunohistochemistry has not reached its limits by far and TAT is an important progressive step in this developmental process.
von Wasielewski
R
R
Pathologisches Institut, Medizinischen Hochschule Hannover, Germany.
Mengel
M
M
Gignac
S
S
Wilkens
L
L
Werner
M
M
Georgii
A
A
D000911:Antibodies, Monoclonal; D001710:Biotin / Q000031:analogs & derivatives*; D005069:Evaluation Studies as Topic; D007150:Immunohistochemistry / Q000379:methods*; D012680:Sensitivity and Specificity; D014443:Tyrosine / Q000031:analogs & derivatives*
D000911:Antibodies, Monoclonal; D014443:Tyrosine; D001710:Biotin