iFluor® 350 Tyramide

iFluor® 350 Tyramide is a blue-fluorescent signal amplification reagent for sensitive detection of low-abundance targets in cells and tissues using horseradish peroxidase (HRP)-mediated labeling.

Tyramide Signal Amplification: Enhances detection sensitivity through HRP-catalyzed fluorescence signal amplification.
Spectral Properties: Excitation at 345 nm and emission at 450 nm.
HRP-Mediated Labeling: Enables localized covalent deposition of fluorescent labels near target sites.
Filter Compatibility: Supports fluorescence detection using standard DAPI filter sets.
Applications: Suitable for amplified immunofluorescence and immunohistochemical staining of low-abundance targets.

Chemical structure for iFluor® 350 Tyramide.

Protocol

SDS

Catalog	Size	Price	Quantity
45095	200 Slides	Price

Overview

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® 350 tyramide contains the bright iFluor® 350 that can be readily detected with the standard DAPI filter set. It is an excellent replacement for Alexa Fluor® 350 tyramide (Alexa Fluor® is the trademark of ThermoFisher) or other comparable fluorescent tyramide conjugates.

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of iFluor® 350 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	231.241 µL	1.156 mL	2.312 mL	11.562 mL	23.124 mL
5 mM	46.248 µL	231.241 µL	462.481 µL	2.312 mL	4.625 mL
10 mM	23.124 µL	115.62 µL	231.241 µL	1.156 mL	2.312 mL

Molarity calculator

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

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
mg	÷	432.45 g/mol	=	mL	x	mM	=	mmol

References

View all 11 references: Citation Explorer

Ultrastructure of light-activated axons following optogenetic stimulation to produce late-phase long-term potentiation.

Authors:

Kuwajima, Masaaki and Ostrovskaya, Olga I and Cao, Guan and Weisberg, Seth A and Harris, Kristen M and Zemelman, Boris V

Journal:

PloS one (2020): e0226797

Intensive Immunofluorescence Staining Methods for Low Expression Protein: Detection of Intestinal Stem Cell Marker LGR5.

Authors:

Yamazaki, Masaki and Kato, Atsuhiko and Zaitsu, Yoko and Watanabe, Takeshi and Iimori, Makoto and Funahashi, Shinichi and Kitao, Hiroyuki and Saeki, Hiroshi and Oki, Eiji and Suzuki, Masami

Journal:

Acta histochemica et cytochemica (2015): 159-64

Tyramide signal amplification for analysis of kinase activity by intracellular flow cytometry.

Authors:

Clutter, Matthew R and Heffner, Garrett C and Krutzik, Peter O and Sachen, Kacey L and Nolan, Garry P

Journal:

Cytometry. Part A : the journal of the International Society for Analytical Cytology (2010): 1020-31

Methoxychlor and estradiol induce oxidative stress DNA damage in the mouse ovarian surface epithelium.

Authors:

Symonds, Daniel A and Merchenthaler, Istvan and Flaws, Jodi A

Journal:

Toxicological sciences : an official journal of the Society of Toxicology (2008): 182-7

Genotyping of phenotypically defined cells in neoplasia: enhanced immunoFISH via tyramide signal amplification (TSA) segregates immunophenotypically-defined cell populations for gated genotyping.

Authors:

Tubbs, Raymond R and Das, Kingshuk and Cook, James R and Pettay, James D and Roche, Patrick C and Grogan, Thomas

Journal:

Journal of molecular histology (2007): 129-34

Physical properties

Molecular weight	432.45
Solvent	DMSO

Spectral properties

Correction factor (260 nm)	0.83
Correction factor (280 nm)	0.23
Extinction coefficient (cm ^-1 M ^-1)	20000 ¹
Excitation (nm)	345
Emission (nm)	450
Quantum yield	0.95 ¹

Usage and storage

Intended use	Research Use Only (RUO)
Storage	Freeze (< -15 °C); Minimize light exposure

Instrument settings

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

Telephone
Fax
Email	sales@aatbio.com
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Bulk request	Inquire
Custom size	Inquire
Technical support	Contact us
Request quotation	Request
Purchase order	Send to sales@aatbio.com
Shipping	Standard overnight for United States, inquire for international

Page updated on November 4, 2026