Name: iFluor® 546 Tyramide
Brand: AAT Bioquest
SKU: 45103
Price: 332 USD
Availability: InStock

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.

Protocol

SDS

COA

Catalog	Size	Price	Quantity
45103	200 slides	Price

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of iFluor® 546 Tyramide to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	77.95 µL	389.748 µL	779.496 µL	3.897 mL	7.795 mL
5 mM	15.59 µL	77.95 µL	155.899 µL	779.496 µL	1.559 mL
10 mM	7.795 µL	38.975 µL	77.95 µL	389.748 µL	779.496 µL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
mg	÷	1282.88 g/mol	=	mL	x	mM	=	mmol

References

View all 50 references: Citation Explorer

Microwaving and Fluorophore-Tyramide for Multiplex Immunostaining on Mouse Adrenals - Using Unconjugated Primary Antibodies from the Same Host Species.

Authors:

Lyu, Qiongxia and Zheng, Huifei Sophia and Laprocina, Karly and Huang, Chen-Che Jeff

Journal:

Journal of visualized experiments : JoVE (2020)

Detection of Cytokine Receptors Using Tyramide Signal Amplification for Immunofluorescence.

Authors:

Wang, Herui and Pangilinan, Ryan L and Zhu, Yan

Journal:

Methods in molecular biology (Clifton, N.J.) (2020): 89-97

Procedural Requirements and Recommendations for Multiplex Immunofluorescence Tyramide Signal Amplification Assays to Support Translational Oncology Studies.

Authors:

Parra, Edwin Roger and Jiang, Mei and Solis, Luisa and Mino, Barbara and Laberiano, Caddie and Hernandez, Sharia and Gite, Swati and Verma, Anuj and Tetzlaff, Michael and Haymaker, Cara and Tamegnon, Auriole and Rodriguez-Canales, Jaime and Hoyd, Clifford and Bernachez, Chantale and Wistuba, Ignacio

Journal:

Cancers (2020)

Sensitive Multiplexed Fluorescent In Situ Hybridization Using Enhanced Tyramide Signal Amplification and Its Combination with Immunofluorescent Protein Visualization in Zebrafish.

Authors:

Lauter, Gilbert and Söll, Iris and Hauptmann, Giselbert

Journal:

Methods in molecular biology (Clifton, N.J.) (2020): 397-409

Optimization of prostate cancer cell detection using multiplex tyramide signal amplification.

Authors:

Roy, Sounak and Axelrod, Haley D and Valkenburg, Kenneth C and Amend, Sarah and Pienta, Kenneth J

Journal:

Journal of cellular biochemistry (2019): 4804-4812

Physical properties

Molecular weight	1282.88
Solvent	DMSO

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-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12171501

Instrument settings

Fluorescence microscope
Excitation	Cy3/TRITC filter set
Emission	Cy3/TRITC filter set
Recommended plate	Black wall/clear bottom

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Page updated on October 16, 2025