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iFluor® 546 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® 546 tyramide contains the bright iFluor® 546 that can be readily detected with the standard TRITC or Cy3 filter set. iFluor® dyes have higher fluorescence intensity, increased photostability, and enhanced water solubility, resulting in fluorescence signals with significantly higher precision and sensitivity. iFluor® 546 is an excellent replacement for Alexa Fluor® 546 tyramide (Alexa Fluor® is the trademark of ThermoFisher), TRITC tyramide, or other comparable fluorescent tyramide conjugates.
Microtubules of fixed HeLa cells were labeled with anti-α tubulin mouse mAb followed by HRP-labeled goat anti-mouse IgG (Cat No. 16728). The fluorescence signal was developed using Alexa Fluor® 546 tyramide or iFluor® 546 tyramide (Cat No. 45103) and detected with a TRITC/Cy3 filter set. iFluor® 546 tyramide shows significantly higher fluorescence intensity than Alexa Fluor® 546 tyramide under the same conditions.
Microtubules of fixed HeLa cells were labeled with anti-α tubulin mouse mAb followed by HRP-labeled goat anti-mouse IgG (Cat No. 16728). The fluorescence signal was developed using Alexa Fluor® 546 tyramide or iFluor® 546 tyramide (Cat No. 45103) and detected with a TRITC/Cy3 filter set. iFluor® 546 tyramide shows significantly higher fluorescence intensity than Alexa Fluor® 546 tyramide under the same conditions.
CatalogSize
Price
Quantity
45103200 slides
Price
 
Physical properties

Molecular weight1282.88
SolventDMSO
Spectral properties

Correction factor (260 nm)0.25
Correction factor (280 nm)0.15
Extinction coefficient (cm -1 M -1)
100000
1
Excitation (nm)541
Emission (nm)557
Quantum yield
0.67
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
UNSPSC12171501
Instrument settings

Fluorescence microscope
ExcitationCy3/TRITC filter set
EmissionCy3/TRITC filter set
Recommended plateBlack wall/clear bottom
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Page updated on October 11, 2025