Cy3 tyramide

For many immunohistochemical (IHC) applications, the traditional enzymatic amplification procedures are sufficient for achieving adequate antigen detection. However, several factors limit the sensitivity and utility of these procedures. 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 translates ultrasensitive detection of low-abundance targets and the use of smaller amounts of antibodies and hybridization probes. In immunohistochemical applications, sensitivity enhancements derived from TSA method allow primary antibody dilutions to be increased to reduce nonspecific background signals, and can overcome weak immunolabeling caused by suboptimal fixation procedures or low levels of target expression. Cy3 tyramide contains the bright Cy3 that can be readily detected with the standard Cy3 filter set.

<p>Immunofluorescent image of paraffin-embedded human lung carcinoma labeled with EpCAM Rabbit mAb followed with HRP-labeled goat anti-rabbit IgG (H+L) (Cat#16793). The signal was developed with AAT’s Cy3 tyramide (Cat#11065, Red). Cells were also counterstained with DAPI (Blue).</p>

Protocol

SDS

COA

Catalog	Size	Price	Quantity
11065	1 mg	Price

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of Cy3 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	115.746 µL	578.73 µL	1.157 mL	5.787 mL	11.575 mL
5 mM	23.149 µL	115.746 µL	231.492 µL	1.157 mL	2.315 mL
10 mM	11.575 µL	57.873 µL	115.746 µL	578.73 µL	1.157 mL

Molarity calculator

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

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
mg	÷	863.96 g/mol	=	mL	x	mM	=	mmol

Citations

View all 12 citations: Citation Explorer

Association of alterations in transcriptomics and intestinal immune responses with Bifidobacterium longum BAA2573 in improving dextran sulfate sodium-induced colitis

Authors:

Hao, Wujuan and Gu, Lan and Zhou, Renmin and Huang, Cuilan and Wang, Xuyang and Liu, Yanshan and Lin, Qiong

Journal:

BMC Gastroenterology (2025): 1--15

Immunostimulatory DNA Tetrahedron-Based Nanovaccine Combined With Immune Checkpoint PD-1 Blockade for Boosting Systemic Immune Responses Against Oral Squamous Cell Carcinoma

Authors:

Yang, Xueting and Wang, Yun and Yang, Yan and Zhang, Sicheng and Wang, Dianri and Luo, Yi and Shui, Chunyan and Cai, Yongcong and Yang, Ruoyi and Dong, Shuang and others,

Journal:

Aggregate (2025): e70061

Anti-TGF-$\beta$/PD-L1 bispecific antibody synergizes with radiotherapy to enhance antitumor immunity and mitigate radiation-induced pulmonary fibrosis

Authors:

Wu, Yuze and Yan, Yuheng and Guo, Yarong and Niu, Mengke and Zhou, Binghan and Zhang, Jing and Zhou, Pengfei and Chu, Qian and Mei, Qi and Yi, Ming and others,

Journal:

Journal of Hematology \& Oncology (2025): 1--20

A novel vimentin variant from tumor-associated macrophages directs cancer metastasis by engaging IGF-1R

Authors:

Jiu, Yaming and Miao, Chenglin

Journal:

bioRxiv (2025): 2025--02

Allopregnanolone relieves paclitaxel induced mechanical hypersensitivity via inhibiting spinal cord PGE2-EP2 mediated microglia-neuron signaling

Authors:

Guo, Kunlin and Gao, Lei and Li, Ping and Feng, Shanwu and Zhao, Liping and Wang, Xian

Journal:

IBRO Neuroscience Reports (2025)

References

View all 74 references: Citation Explorer

Tyramide Signal Amplification for Immunofluorescent Enhancement

Authors:

Faget L, Hnasko TS.

Journal:

Methods Mol Biol (2015): 161

Enhanced detection of Porcine reproductive and respiratory syndrome virus in fixed tissues by in situ hybridization following tyramide signal amplification

Authors:

Trang NT, Hirai T, Ngan PH, Lan NT, Fuke N, Toyama K, Yamamoto T, Yamaguchi R.

Journal:

J Vet Diagn Invest (2015): 326

Rapid and sensitive detection of Escherichia coli O157:H7 in milk and ground beef using magnetic bead-based immunoassay coupled with tyramide signal amplification

Authors:

Aydin M, Herzig GP, Jeong KC, Dunigan S, Shah P, Ahn S.

Journal:

J Food Prot (2014): 100

Multiplexed immunohistochemistry, imaging, and quantitation: a review, with an assessment of Tyramide signal amplification, multispectral imaging and multiplex analysis

Authors:

Stack EC, Wang C, Roman KA, Hoyt CC.

Journal:

Methods (2014): 46

KSHV cell attachment sites revealed by ultra sensitive tyramide signal amplification (TSA) localize to membrane microdomains that are up-regulated on mitotic cells

Authors:

Garrigues HJ, Rubinchikova YE, Rose TM.

Journal:

Virology (2014): 75

Physical properties

Molecular weight	863.96
Solvent	DMSO

Spectral properties

Correction factor (260 nm)	0.07
Correction factor (280 nm)	0.073
Extinction coefficient (cm ^-1 M ^-1)	150000 ¹
Excitation (nm)	555
Emission (nm)	569
Quantum yield	0.15 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 September 20, 2025