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What is Tyramide Signal Amplification (TSA)?

Posted March 10, 2020


Answer

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Tyramide signal amplification (TSA) is a novel technique designed to enhance the detection of low-abundance targets (e.g. proteins and nucleic acid sequences). It can be readily integrated into any application that utilizes horseradish peroxidase (HRP) in its protocol. These applications include ELISA, immunocytochemistry (ICC), immunohistochemistry (IHC) and in situ hybridization (FISH). Advantages of tyramide signal amplification, include:

  • Detection of low-abundance targets
  • Higher precision and sensitivity up to 100-fold that of conventional ICC, IHC and FISH methods
  • Easy integration with ICC, IHC and FISH applications
  • Conservation of primary and secondary antibodies

TSA Principle & Workflow

TSA - sometimes called Catalyzed Reporter Deposition or CARD, is an enyzme-mediated detection method that utilizes the catalytic activity of HRP for the covalent deposition of labeled tyramide on and near target proteins or nucleic acid sequences in situ. In the presence of hydrogen peroxide (H2O2), HRP converts labeled tyramide substrates into highly-reactive, short-lived tyramide radicals that rapidly bind to tyrosine residues on and proximal to the enzyme site. These labels can be detected by standard chromogenic or fluorescent techniques.

TSA labeling procedure consists of the following steps:

  1. Sample preparation – fixation and permeabilization of cells or tissue sample
  2. Blocking of nonspecific epitopes
  3. Binding with primary antibody, biotinylated primary antibody or biotinylated nucleic acid probe
  4. Binding with HRP-secondary antibody or HRP-streptavidin
  5. Incubation with tyramide substrate
  6. Mount sample and image signal

 

TSA workflow illustration
Figure 1. Workflow for tyramide signal amplification (TSA). With workflow similar to conventional ICC and IHC methods, TSA can achieve sensitive detection of desired targets in a few simple steps.

Available tyramide reagents

Product Name Ex/Em (nm) ε Φ Size Cat#
Azido-Cy5 tyramide 650/669 250,000 0.27 1 mg 11061
Cy3 tyramide 554/568 150,000 0.04 1 mg 11065
Cy5 tyramide 650/669 250,000 0.27 1 mg 11066
AF488 tyramide reagent 499/520 73,000 0.92 200 slides 11070
AF546 tyramide reagent 561/572 112,000 0.1 200 slides 11075
AF594 tyramide reagent 590/618 92,000 0.66 200 slides 11082
iFluor® 488 tyramide 491/516 75,000 0.9 200 slides 45100
iFluor® 555 Tyramide 556/569 100,000 0.64 200 Slides 45105
iFluor® 647 Tyramide 654/669 250,000 0.25 200 Slides 45110
Additional resources

iFluor™ 488 tyramide Product Page

Power Styramide™ Signal Amplification (PSA™)