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iFluor® 647 Tyramide
For many immunohistochemical (IHC) applications, traditional enzymatic amplification procedures are sufficient for achieving adequate antigen detection. However, several factors limit their sensitivity and utility. Tyramide signal amplification (TSA) has proven to be a particularly versatile and powerful enzyme amplification technique with improved assay sensitivity. TSA is based on the ability of HRP, in the presence of low concentrations of hydrogen peroxide, to convert labeled tyramine-containing substrate into an oxidized, highly reactive free radical that can covalently bind to tyrosine residues at or near the HRP. To achieve maximal IHC detection, tyramine is prelabeled with a fluorophore. The signal amplification conferred by the turnover of multiple tyramide substrates per peroxidase label results in the ability to detect low-abundance targets with ultrasensitive precision and reduces the amount of antibodies and hybridization probes needed. In IHC applications, this method can also enhance sensitivity in cases where the primary antibody dilution needs to be increased to reduce nonspecific background signals or overcome weak immunolabeling due to suboptimal fixation procedures or low levels of target expression. The iFluor® 647 tyramide contains the bright iFluor® 647 that can be readily detected with the standard Cy5 filter set. It is an excellent replacement for Alexa Fluor® 647 tyramide (Alexa Fluor® is the trademark of ThermoFisher), Cy5 tyramide, or other comparable fluorescent tyramide conjugates.
Formalin-fixed, paraffin-embedded (FFPE) human lung tissue was labeled with anti-EpCAM mouse mAb followed by HRP-labeled goat anti-mouse IgG (Cat No. 16728). The fluorescence signal was developed using iFluor® 647 tyramide (Cat No. 45110) and detected with a Cy5 filter set. Nuclei (blue) were counterstained with DAPI (Cat No. 17507).
Formalin-fixed, paraffin-embedded (FFPE) human lung tissue was labeled with anti-EpCAM mouse mAb followed by HRP-labeled goat anti-mouse IgG (Cat No. 16728). The fluorescence signal was developed using iFluor® 647 tyramide (Cat No. 45110) and detected with a Cy5 filter set. Nuclei (blue) were counterstained with DAPI (Cat No. 17507).
protocol
Protocol
sds
SDS
COA
CatalogSize
Price
Quantity
45110200 Slides
Price
 
Physical properties
Molecular weight1092.20
SolventDMSO
Spectral properties
Correction factor (260 nm)0.03
Correction factor (280 nm)0.03
Correction factor (656 nm)0.0793
Extinction coefficient (cm -1 M -1)
250000
1
Excitation (nm)656
Emission (nm)670
Quantum yield
0.25
1
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12352200
Instrument settings
Fluorescence microscope
ExcitationCy5 filter set
EmissionCy5 filter set
Recommended plateBlack wall/clear bottom
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Page updated on October 10, 2025