Signal amplification in immunohistochemistry: loose-jointed deformable heteropolymeric HRP conjugates vs. linear polymer backbone HRP conjugates.
Improvements in reagents and protocols for immunohistochemistry have led to increased sensitivity of detection systems. A significant level of signal amplification was achieved by the chain-polymer conjugate technology utilizing enzyme-labeled inert "backbone" molecule of dextran (Dako). However, the relatively large size of the dextran molecule in aqueous phase appears to create spatial hindrance compromising the penetrative ability of the detection reagent. Novel AmpliStain™ detection systems (SDT GmbH, Baesweiler, Germany) seem to overcome these constraints offering a more compact and deformable conjugate design that facilitates agile penetration through the narrowest diffusion pathways in tissue sections. Here, we compared the level of signal amplification achievable with AmpliStain™-HRP (SDT) and EnVision™+-HRP (Dako). Our results show that the AmpliStain™-HRP systems allow higher dilutions of primary antibodies in both immunohistochemistry and ELISA. Compared with EnVision™+, anti-mouse AmpliStain™ enables at least three times more sensitive detection of mouse antibodies, whereas anti-rabbit AmpliStain™ is ten times more sensitive than anti-rabbit EnVision™+.
Institute for Hematopathology, Fangdieckstr. 75a, 22547 Hamburg, Germany. buchwalow@pathologie-hh.de
Igor
I
Buchwalow
Werner
W
Boecker
Eduard
E
Wolf
Vera
V
Samoilova
Markus
M
Tiemann
D000328:Adult; D000368:Aged; D000818:Animals; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology; D003911:Dextrans / Q000737:chemistry / Q000378:metabolism*; D005260:Female; D006735:Horseradish Peroxidase / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D051379:Mice; D008875:Middle Aged; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D014066:Palatine Tonsil / Q000378:metabolism / Q000473:pathology; D011108:Polymers; D011817:Rabbits; D012680:Sensitivity and Specificity; D015398:Signal Transduction; D012867:Skin / Q000378:metabolism / Q000473:pathology; D055815:Young Adult
D003911:Dextrans; D011108:Polymers; D006735:Horseradish Peroxidase
Antibodies; Detection systems; Differential diagnosis; Immunohistochemistry; Nuclear antigens; Signal amplification