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Highly Sensitive Tyramide Signal Amplification Kits

Highly Sensitive Tyramide Signal Amplification Kits

Cambridge Bioscience offers highly sensitive kits for detecting low-abundance targets in fluorescent immunocytochemistry (ICC), immunohistochemistry (IHC) and in situ hybridisation (FISH). With sensitivity up to 100 times higher than conventional methods and requiring less antibody, these tyramide signal amplification kits contain the bright and photostable CF® dyes or biotin with anti-mouse, anti-rabbit or HRP streptavidin conjugates available.

Tyramide signal amplification

Tyramide signal amplification

Horseradish peroxidase (HRP) catalyses the covalent reaction of tyramide to tyrosine residues. In the tyramide signal amplification technique, immunostaining is performed with HRP-conjugated antibodies or streptavidin. Subsequent reaction with fluorescent tyramide causes large numbers of fluorescent dye molecules to be deposited in the vicinity of the antibody, generating dramatically stronger signal compared to conventional immunofluorescence.

Tyramide signal amplification protocol

Benefits Of Using These Tyramide Signal Amplification Kits
• 100-fold greater sensitivity than conventional methods
• High signal-to-background ratio
• All critical reagents for tyramide labelling included
• Use less antibody
• Choice of biotin tyramide or one of six CF® dye tyramides (CF®488A, CF®543, CF®568, CF®594, CF®640R, or CF®680R)
• Choice of HRP conjugate (goat anti-mouse, goat anti-rabbit or streptavidin-HRP)
• Simplified primary antibody panel design for multiplex IHC

Multiplex tyramide labeling of FFPE tissues

Multiplex tyramide labelling of FFPE tissues. CF®488A-tyramide labelling pan-CK (green); Cy®3-tyramide labelling histone H1 (red); CF®640R-tyramide labeling ZO1 (magenta). Primary antibodies were from mouse & secondary antibody was HRP-conjugated goat anti-mouse. Each labelling was performed sequentially, with antibody removal by microwave treatment.

Sequential labelling of formaldehyde-fixed HeLa cells with two Tyramide Amplification Kits

Sequential labelling of formaldehyde-fixed HeLa cells with two Tyramide amplification kits. Mitochondria (green) visualised with rabbit anti-COXIV primary antibody & tyramide amplification kit with HRP goat anti-rabbit IgG & CF®488A-tyramide, followed by peroxidase quenching. Nucleoli (magenta) visualised with mouse anti-cyclin B1 primary antibody & tyramide amplification kit with HRP Goat anti-mouse IgG & CF®568-tyramide. Actin (red) detected with CF®640R-phalloidin & cell nuclei (blue) stained with DAPI.

Applications
• Fluorescent immunocytochemistry (ICC)
• Immunohistochemistry (IHC)
• In situ hybridisation (FISH)

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