Tyramide Signal Amplification

Fluorescent Dye Tyramides

A wide selection of bright and photostable fluorescent CF® Dye tyramides. Biotin and DNP conjugates also available.

Tyramide Signal Amplification Kits

All-in-one kits that contain all critical reagents for tyramide labeling of cells or tissue, available with CF® Dyes or biotin.

Compatible with ICC, IHC and FISH

Similar workflow to conventional ICC, IHC and FISH; and compatible with multiplex fluorescent detection

Boost Your Signal Up to 100-Fold

Tyramide signal amplification (TSA), sometimes referred to as Catalyzed Reporter Deposition (CARD), is a highly sensitive method capable of boosting signal up to 100-fold over conventional methods. This enables the detection of low-abundance targets for fluorescent immunocytochemistry (ICC), immunohistochemistry (IHC), and in situ hybridization (FISH) applications.

How TSA Works

TSA uses horseradish peroxidase (HRP) to catalyze deposition of labeled tyramide near a target protein or nucleic acid. In low concentrations of H₂O₂, HRP activates tyramide to covalently bind tyrosine residues, producing high-density labeling near the epitope. This enables exceptional sensitivity. Tyramide can be labeled with fluorophores or haptens like biotin or DNP.

Illustration of the tyramide signal amplification system. A cell or tissue sample is labeled with primary and secondary antibody using conventional methods. The... See More

Example of serial tyramide signal amplification workflow for multiplex detection.

Multiple rounds of tyramide signal amplification can be performed for multicolor detection (see our Tech Tip: Multicolor Fluorescence Imaging using Tyramide Amplification Kits). When TSA followed by antibody removal is used for multiplex fluorescent IHC, TSA not only facilitates detection of low-abundance targets, but also simplifies antibody panel design because primary antibodies of choice may be used, irrespective of host species or isotype.

Advantages of Tyramide Signal Amplification

Detect low-abundance targets
Compatible with ICC, IHC and FISH
Sensitivity up to 100-fold that of conventional methods
Similar workflow to conventional ICC, IHC and FISH
Use less primary antibody
Simplified primary antibody panel design for multiplex IHC

Tyramide Signal Amplification Kits

Our kits provide all critical reagents for tyramide labeling
Choose your tyramide: biotin tyramide or one of six CF® Dye tyramides
Choose your HRP conjugate: goat anti-mouse, goat anti-rabbit, or streptavidin
Kits also include Amplification Buffer and BSA for blocking

Tyramide Amplification Buffers

Tyramide Amplification Buffer Plus has enhanced sensitivity for TSA, with improved brightness, specificity, and sensitivity over our original buffer.
Ready-to-Use Tyramide Amplification Buffer was our original buffer, and has been replaced with the above buffer. The advantage of this buffer is that there is no need to add hydrogen peroxide.

CF® Dye Tyramides — Multiplex tyramide labeling of FFPE tissues. Cytokeratin (pan) was labeled with CF®488A-tyramide (green); Histone H1 was labeled with Cy®3-tyramide (red); ZO1 w... See More

Tyramide Signal Amplification Kits

Tyramide Label	Ex/Em	Secondary conjugate	Catalog no.
CF®488A	490/515 nm	Goat anti-mouse HRP	33000
		Goat anti-rabbit HRP	33001
		Streptavidin HRP	33002
CF®543	541/560 nm	Goat anti-mouse HRP	33003
		Goat anti-rabbit HRP	33004
		Streptavidin HRP	33005
CF®568	562/583 nm	Goat anti-mouse HRP	33006
		Goat anti-rabbit HRP	33007
		Streptavidin HRP	33008
CF®594	593/614 nm	Goat anti-mouse HRP	33009
		Goat anti-rabbit HRP	33010
		Streptavidin HRP	33011
CF®640R	642/662 nm	Goat anti-mouse HRP	33012
		Goat anti-rabbit HRP	33013
		Streptavidin HRP	33014
CF®680R	680/701 nm	Goat anti-mouse HRP	33015
		Goat anti-rabbit HRP	33016
		Streptavidin HRP	33017
Biotin-XX	N/A	Goat anti-mouse HRP	33018
		Goat anti-rabbit HRP	33019
		Streptavidin HRP	33020

Standalone Dye and Hapten Labeled Tyramides

Tyramide label	Ex/Em	Size	Catalog no.
CF®350	347/448 nm	0.5 mg	92170
CF®405L	395/545 nm	0.5 mg	92198
CF®405S	404/431 nm	0.5 mg	92197
CF®405M	408/452 nm	0.5 mg	96057
CF®430	426/498 nm	0.5 mg	96053
CF®488A	490/515 nm	0.5 mg	92171
FITC	492/514 nm	0.5 mg	96018
CF®514	516/548 nm	0.5 mg	92199
CF®532	527/558 nm	0.5 mg	96066
CF®543	541/560 nm	0.5 mg	92172
CF®550R	551/577 nm	0.5 mg	96077
CF®555	555/565 nm	0.5 mg	96021
Cyanine 555	555/565 nm	0.5 mg	96020
CF®568	562/583 nm	0.5 mg	92173
CF®583R	586/609 nm	0.5 mg	96085
CF®594	593/614 nm	0.5 mg	92174
CF®620R	617/639 nm	0.5 mg	92194
CF®640R	642/662 nm	0.5 mg	92175
CF®647	650/665 nm	0.5 mg	96022
CF®660R	663/682 nm	0.5 mg	92195
CF®680R	680/701 nm	0.5 mg	92196
CF®710	712/736 nm	0.5 mg	96127
CF®725	729/755 nm	0.5 mg	96128
CF®740	742/767 nm	0.5 mg	96124
CF®750	755/779 nm	0.5 mg	96052
CF®754	748/793 nm	0.5 mg	96090
Biotin-XX	N/A	0.5 mg	92176
DNP	N/A	0.5 mg	96019

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Fluorescence Spectra Viewer

Compare and combine dyes, lasers, and emission filters.

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CF® Dye Selection Tool

Find the optimal CF® Dye for specific laser/filter combinations or different applications.

Blog

Seeing More and Brighter Colors with Tyramide Signal Amplification

Tech Tip

Tech Tip: Multi-Color Fluorescence Imaging Using Biotium’s Tyramide Amplification Kits

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FAQs

Do you offer alternatives to Akoya Opal™ tyramides?

Biotium developed TyraMax™ Amplification Dyes as high-performance TSA dyes, offering brighter, more photostable signals than Aluora® and Opal® reagents. They also remain stable in amplification buffer for up to 24 hours, facilitating automated staining workflows. TyraMax™ Dyes are offered as standalone dye solutions, as 3-color or 5-color dye sets plus DAPI counterstain, and in a sampler for custom panel optimization. See below for alternatives to specific Aluora® and Opal® reagents. Learn more about our reagents for tyramide signal amplification (TSA).

TyraMax™ Amplification Dyes

Dye	Abs/Em (nm)	Laser Line	Detection channel	Dye Features	Size	Catalog No.
TyraMax™ 410/450	408/452	405 nm	DAPI/ Alexa Fluor® 405	Unique tyramide color, spectrally similar to Alexa Fluor® 405	20 uL	96134-20UL
TyraMax™ 410/450	408/452	405 nm	DAPI/ Alexa Fluor® 405		100 uL	96134-100UL
TyraMax™ 430/500	421/497	405 nm	FITC	Brighter than Aluora® 430 and Opal® 480	20 uL	96135-20UL
TyraMax™ 430/500	421/497	405 nm	FITC	Brighter than Aluora® 430 and Opal® 480	100 uL	96135-100UL
TyraMax™ 400/550	394/547	405 nm	FITC	Unique tyramide color, spectrally similar to Pacific Orange®	20 uL	96136-20UL
TyraMax™ 400/550	394/547	405 nm	FITC		100 uL	96136-100UL
TyraMax™ 490/520	497/518	488 nm	FITC	Brighter than Opal® 520, replacement for Aluora® 488	20 uL	96137-20UL
TyraMax™ 490/520	497/518	488 nm	FITC	Brighter than Opal® 520, replacement for Aluora® 488	100 uL	96137-100UL
TyraMax™ 555/565	594/569	555 nm or 561 nm	TRITC	Brighter than Opal® 570, replacement for Aluora® 555	20 uL	96138-20UL
TyraMax™ 555/565	594/569	555 nm or 561 nm	TRITC	Brighter than Opal® 570, replacement for Aluora® 555	100 uL	96138-100UL
TyraMax™ 560/580	562/584	555 nm or 561 nm	TRITC	Alternative to Aluora® 555, Opal® 570 with superior photostability	20 uL	96139-20UL
TyraMax™ 560/580	562/584	555 nm or 561 nm	TRITC		100 uL	96139-100UL
TyraMax™ 630/650	631/650	633 nm or 640 nm	Cy®5	Bright and photostable alternative to Aluora® 647, Opal® 650	20 uL	96140-20UL
TyraMax™ 630/650	631/650	633 nm or 640 nm	Cy®5		100 uL	96140-100UL
TyraMax™ 647/670	650/670	633 nm or 640 nm	Cy®5	Brighter than Opal® 650, replacement for Aluora® 647	20 uL	96141-20UL
TyraMax™ 647/670	650/670	633 nm or 640 nm	Cy®5	Brighter than Opal® 650, replacement for Aluora® 647	100 uL	96141-100UL
TyraMax™ 660/680	663/683	633 nm or 640 nm	Alexa Fluor® 680	Unique tyramide color, spectrally similar to Alexa Fluor® 660. Brighter and more photostable Opal® 690 when excited at 640 nm	20 uL	96142-20UL
TyraMax™ 660/680	663/683	633 nm or 640 nm	Alexa Fluor® 680		100 uL	96142-100UL
TyraMax™ 680/700	680/701	685 nm (detectable with 640 nm excitation)	Alexa Fluor® 680	Brighter and more photostable than Opal® 690 when excited at 685 nm	20 uL	96143-20UL
TyraMax™ 680/700	680/701	685 nm (detectable with 640 nm excitation)	Alexa Fluor® 680		100 uL	96143-100UL
TyraMax™ 710/740	711/736	685 nm	Alexa Fluor® 700	Brighter and more photostable than Aluora® 700	20 uL	96144-20UL
TyraMax™ 710/740	711/736	685 nm	Alexa Fluor® 700	Brighter and more photostable than Aluora® 700	100 uL	96144-100UL
TyraMax™ 740/770	742/768	730 nm	Alexa Fluor® 750	Single-step detection, unlike Opal® 780. Stable in amplification buffer for up to 24 hours, unlike Aluora® 750	20 uL	96145-20UL
TyraMax™ 740/770	742/768	730 nm	Alexa Fluor® 750		100 uL	96145-100UL

TyraMax™ 3-Plex Amplification Dye Set with DAPI (Cat. No. 33029)

Component #	Component Name	Size
96137-100UL	TyraMax™ 490/520 Amplification Dye, 100X, 100 uL	For 100 samples using 100 uL staining volume
96139-100UL	TyraMax™ 560/580 Amplification Dye, 100X, 100 uL
96140-100UL	TyraMax™ 630/650 Amplification Dye, 100X, 100 uL
99897-50UL	DAPI, 1000X, 50 uL

TyraMax™ 5-Plex Amplification Dye Set with DAPI (Cat. No. 33030)

Component #	Component Name	Size
96137-100UL	TyraMax™ 490/520 Amplification Dye, 100X, 100 uL	For 100 samples using 100 uL staining volume
96139-100UL	TyraMax™ 560/580 Amplification Dye, 100X, 100 uL
96140-100UL	TyraMax™ 630/650 Amplification Dye, 100X, 100 uL
96143-100UL	TyraMax™ 680/700 Amplification Dye, 100X, 100 uL
96145-100UL	TyraMax™ 740/770 Amplification Dye, 100X, 100 uL
99897-50UL	DAPI, 1000X, 50 uL

TyraMax™ Amplification Dye Spectral Sampler (Cat. No. 33031)

Component #	Component Name	Size
96134-20UL	TyraMax™ 410/450	For 20 samples per vial using 100 uL staining volume
96135-20UL	TyraMax™ 430/500
96136-20UL	TyraMax™ 400/550
96137-20UL	TyraMax™ 490/520
96138-20UL	TyraMax™ 555/565
96139-20UL	TyraMax™ 560/580
96140-20UL	TyraMax™ 630/650
96141-20UL	TyraMax™ 647/670
96142-20UL	TyraMax™ 660/680
96143-20UL	TyraMax™ 680/700
96144-20UL	TyraMax™ 710/740
96145-20UL	TyraMax™ 740/770

Aluora is a registered trademark of Thermo Fisher Scientific; Opal is a registered trademark of Akoya Biosciences.

Can I incubate multiple primary antibodies combined with tyramide amplificaton?

Our Tyramide Amplification Kits have been demonstrated to be robust and versatile for multi-color fluorescence imaging, compatible with dye-labeled antibodies and various cell staining methods (see Figure 1).

To use a Tyramide Amplification Kit in addition to one or more dye-labeled antibodies, follow the kit protocol to fix and block samples; label with primary antibodies; then detect primary antibodies using secondary antibodies. Dye labeled secondary antibodies can be co-incubated with the HRP-conjugated secondary or HRP-streptavidin from the tyramide kit. After washing, perform the CF® Dye tyramide reaction according to the kit protocol. The tyramide reaction does not interfere with the binding of dye-labeled antibodies or other fluorescent staining reagents.

Performing multi-color detection with more than one dye tyramide on the same sample requires sequential tyramide staining reactions, followed by HRP inactivation or antibody stripping between each step. See our tech tip:

Multi-Color Fluorescence Imaging Using Biotium’s Tyramide Amplification Kits

Can the CF® dye tyramides be used with in situ hybridization or RNAscope assays from ACD?

Yes, CF® dye tyramides can be used for in situ hybridization using Tyramide Signal Amplification¹ (TSA). They are also compatible with the RNAscope assays for RISH².

¹ doi: https://doi.org/10.1038/s41598-018-38171-5.

²doi: http://dx.doi.org/10.1101/651083.

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