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Tyramide Signal Amplification

Largest selection of fluorescent dye tyramides
21 bright and photostable fluorescent CF® Dye tyramides. Biotin & DNP conjugates also available
Tyramide Signal Amplification Kits
Providing all critical reagents for tyramide labeling of cells or tissue
Compatible with ICC, IHC and FISH
Similar workflow to conventional ICC, IHC and FISH; and compatible with multiplex fluorescent detection

Tyramide signal amplification (TSA), sometimes called Catalyzed Reporter Deposition (CARD), is a highly sensitive method enabling the detection of low-abundance targets in fluorescent immunocytochemistry (ICC), immunohistochemistry (IHC), and in situ hybridization (FISH) applications.

TSA involves horseradish peroxidase (HRP)-catalyzed deposition of tyramide on and near a target protein or nucleic acid sequence in situ. In the presence of low concentrations of H2O2, HRP is able to convert a labeled tyramide substrate into a highly reactive form that can covalently bind to tyrosine residues on proteins at or near the HRP. This generates high density tyramide labeling and is the reason for the exceptional sensitivity of this system. Tyramide can be labeled with a fluorophore or a hapten (such as biotin or DNP).

Multiple rounds of tyramide signal amplification can be performed for multicolor detection (see Figure 2 and 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 tyramide signal amplification. It has improved brightness, specificity, and sensitivity over our original buffer below.
  • Ready-to-Use Tyramide Amplification Buffer was our original buffer formulation, and has been replaced with the above buffer. The one advantage of this buffer is that there is no need to add hydrogen peroxide.
Figure 1. Illustration of the tyramide signal amplification system. A cell or tissue sample is labeled with primary and secondary antibody using conventional methods. The horseradish peroxidase, conjugated to the secondary antibody, catalyzes the conversion of labeled tyramide into a reactive radical. The tyramide radical then covalently binds to nearby tyrosine residues, providing high-density labeling.
Figure 2. Example of serial tyramide signal amplification workflow for multiplex detection.

Figure 3. 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 was labeled with CF®640R-tyramide (magenta). Primary antibodies were from mouse, and secondary antibody was HRP-conjugated goat anti-mouse. Each labeling was performed sequentially, with antibody removal by microwave treatment between labeling steps.
Figure 4. Sequential labeling of formaldehyde-fixed HeLa cells with two Tyramide Amplification Kits. Mitochondria (green) was labeled with rabbit anti-COXIV primary antibody and Tyramide Amplification Kit with HRP goat anti-rabbit IgG and CF®488A-tyramide, followed by peroxidase quenching. Nucleoli (magenta) visualized with mouse anti-cyclin B1 primary antibody and Tyramide Amplification Kit with HRP Goat anti-mouse IgG and CF®568-tyramide. Actin (red) was detected with CF®640R-phalloidin, and cell nuclei (blue) were stained with DAPI.

Standalone Dye and Hapten Labeled Tyramides

Tyramide label Ex/EmSizeCatalog no.
CF®350347/448 nm0.5 mg 92170
CF®405L395/545 nm0.5 mg92198
CF®405S 404/431 nm0.5 mg92197
CF®405M 408/452 nm0.5 mg96057
CF®430426/498 nm0.5 mg96053
CF®488A490/515 nm0.5 mg92171
FITC492/514 nm0.5 mg96018
CF®514 516/548 nm0.5 mg92199
CF®532 527/558 nm0.5 mg96066
CF®543 541/560 nm0.5 mg92172
CF®550R551/577 nm0.5 mg96077
CF®555 555/565 nm0.5 mg96021
Cyanine 555555/565 nm0.5 mg96020
CF®568 562/583 nm0.5 mg92173
CF®583R586/609 nm0.5 mg96085
CF®594 593/614 nm0.5 mg92174
CF®620R 617/639 nm0.5 mg92194
CF®640R 642/662 nm0.5 mg92175
CF®647 650/665 nm0.5 mg96022
CF®660R 663/682 nm0.5 mg92195
CF®680R 680/701 nm0.5 mg92196
CF®750755/779 nm0.5 mg96052
CF®754748/793 nm0.5 mg96090
Biotin-XX N/A0.5 mg92176
DNP N/A0.5 mg96019

Tyramide Amplification Kit with HRP Goat Anti-Mouse and CF® Dye or Biotin Tyramide

From: $564 Sizes: 1 kitCatalog #:
, 33003, 33006, 33009, 33012, 33015, - 33018View allHide

Tyramide Amplification Kit with HRP Goat Anti-Rabbit and CF® Dye or Biotin Tyramide

From: $564 Sizes: 1 kitCatalog #:
, 33004, 33007, 33010, 33013, 33016, - 33019View allHide

Tyramide Amplification Kit with HRP Streptavidin and CF® Dye or Biotin Tyramide

From: $564 Sizes: 1 kitCatalog #:
, 33005, 33008, 33011, 33014, 33017, - 33020View allHide

Biotin-XX Tyramide

From: $329 Sizes: 0.5 mgCatalog #: 92176

CF® Dye Tyramide

From: $318 Sizes: 0.5 mgCatalog #:
, 96090, 96077, 96066, 92170, 92171, 92172, 92173, 96085, 92174, 92175, 92194, 92195, 92196, 92197, 92198, 92199, 96021, 96022, 96053, - 96057View allHide

Cyanine 555 Tyramide

From: $340 Sizes: 0.5 mgCatalog #: 96020

DNP Tyramide

From: $326 Sizes: 0.5 mgCatalog #: 96019

Tyramide Amplification Buffer Plus

From: $127 Sizes: 250 reactions, 1000 reactionsCatalog #:
, - 22029-TView allHide