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Amplite® Tyramide (TSA) Signal Booster

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 Alexa Fluor™ 488 Tyramide Reagent (Green) with and without Amplite® Tyramide (TSA) Signal Booster. Cells were also counterstained blue with DAPI (Cat#17507).
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 Alexa Fluor™ 488 Tyramide Reagent (Green) with and without Amplite® Tyramide (TSA) Signal Booster. Cells were also counterstained blue with DAPI (Cat#17507).
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 Alexa Fluor™ 488 Tyramide Reagent (Green) with and without Amplite® Tyramide (TSA) Signal Booster. Cells were also counterstained blue with DAPI (Cat#17507).
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Physical properties
SolventDMSO
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure

OverviewpdfSDSpdfProtocol


Tyramide signal amplification (TSA) is a technique used to enhance the signal of target molecules, such as antibodies or nucleic acid probes, in immunohistochemistry and in situ hybridization. This technique is particularly useful when working with samples that have low levels of target molecules or weak signals. TSA works by utilizing the enzymatic activity of horseradish peroxidase (HRP), which can catalyze the deposition of labeled tyramide molecules in close proximity to the target molecules. The tyramide molecules contain a reporter group, such as a fluorescent dye, which generates a detectable signal upon enzymatic reaction. This localized deposition of labeled tyramide molecules leads to signal amplification at the site of the target molecules, making them more easily detectable. There are a number of fluorescent tyramides available for TSA applications. However, some of the commercial tyramide conjugates have low signals. Amplite® Tyramide (TSA) Signal Booster is used to further enhance the TSA signal. It has been validated to enhance the TSA signal of tyramides prepared from Alexa Fluor® 350, 430, 450, 488, 514, 546, 568 and 594, Cy3, rhodamines, fluoresceins and coumarins.

Example protocol


PREPARATION OF STOCK SOLUTIONS

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles

  1. Prepare a stock solution by adding 1 mL of DMSO to the Amplite® Tyramide (TSA) Signal Booster vial.

    Note: Prepare a single unused aliquot of the stock solution and store it at ≤ -20 ºC. Protect it from light and avoid repeated freeze-thaw cycles.

PREPARATION OF WORKING SOLUTION

  1. Add 10 µL of Amplite® Tyramide Signal Booster stock solution to 1 mL of the tyramide working solution.

    Note: For best results, this solution should be used within a few hours of its preparation.

SAMPLE EXPERIMENTAL PROTOCOL

  1. Use the tyramide working solution containing Amplite® Tyramide Signal Booster on tissue samples according to the protocol described by the manufacturer of the tyramide reagent.

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References


View all 50 references: Citation Explorer
Development of an in situ cell-type specific proteome analysis method using antibody-mediated biotinylation.
Authors: Ryu, Taekyung and Kim, Seok-Young and Thuraisamy, Thujitha and Jang, Yura and Na, Chan Hyun
Journal: bioRxiv : the preprint server for biology (2023)
An ultrasensitive electrochemical aptasensor using Tyramide-assisted enzyme multiplication for the detection of Staphylococcus aureus.
Authors: Nguyen, Thi Thanh-Qui and Gu, Man Bock
Journal: Biosensors & bioelectronics (2023): 115199
CRISPR/Cas12a-Triggered Chemiluminescence Enhancement Biosensor for Sensitive Detection of Nucleic Acids by Introducing a Tyramide Signal Amplification Strategy.
Authors: Hu, Tao and Ke, Xinxin and Ou, Yangjing and Lin, Yu
Journal: Analytical chemistry (2022): 8506-8513
Navigating the cellular landscape in tissue: Recent advances in defining the pathogenesis of human disease.
Authors: Chen, Helen Y and Palendira, Umaimainthan and Feng, Carl G
Journal: Computational and structural biotechnology journal (2022): 5256-5263
Detection of hydrocarbon-degrading bacteria on deepwater corals of the northeast Atlantic using CARD-FISH.
Authors: Thompson, Haydn Frank and Gutierrez, Tony
Journal: Journal of microbiological methods (2021): 106277
Phenoxy Radical Reactivity of Nucleic Acids: Practical Implications for Biotinylation.
Authors: Wilbanks, Brandon and Garcia, Brian and Byrne, Shane and Dedon, Peter and Maher, L James
Journal: Chembiochem : a European journal of chemical biology (2021): 1400-1404
Immunohistochemical Detection of 5-Hydroxymethylcytosine and 5-Carboxylcytosine in Sections of Zebrafish Embryos.
Authors: Jessop, Peter and Gering, Martin
Journal: Methods in molecular biology (Clifton, N.J.) (2021): 193-208
Accessibility-dependent topology studies of membrane proteins using a SpyTag/SpyCatcher protein-ligation system.
Authors: Bae, Yoonji and Lee, Sang Kwon and Chae, Young Chan and Park, Chan Young and Kang, Sebyung
Journal: International journal of biological macromolecules (2021): 171-178
Single-cell RNA sequencing of human liver reveals hepatic stellate cell heterogeneity.
Authors: Payen, Valéry L and Lavergne, Arnaud and Alevra Sarika, Niki and Colonval, Megan and Karim, Latifa and Deckers, Manon and Najimi, Mustapha and Coppieters, Wouter and Charloteaux, Benoît and Sokal, Etienne M and El Taghdouini, Adil
Journal: JHEP reports : innovation in hepatology (2021): 100278
Highly Sensitive and Multiplexed In Situ RNA Profiling with Cleavable Fluorescent Tyramide.
Authors: Xiao, Lu and Labaer, Joshua and Guo, Jia
Journal: Cells (2021)