iFluor® 555 Tyramide
Ordering information
Price | |
Catalog Number | |
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
Quotation | Request |
International | See distributors |
Shipping | Standard overnight for United States, inquire for international |
Physical properties
Molecular weight | 1046.41 |
Solvent | DMSO |
Spectral properties
Correction Factor (260 nm) | 0.23 |
Correction Factor (280 nm) | 0.14 |
Extinction coefficient (cm -1 M -1) | 1000001 |
Excitation (nm) | 557 |
Emission (nm) | 570 |
Quantum yield | 0.641 |
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 | 12352200 |
Overview | ![]() ![]() |
See also: Antibodies and Proteomics, Antibody and Protein Labeling, Bioconjugation, Cell Structures and Organelles, Nucleus, DNA and RNA Quantitation, Fluorescent in situ hybridization (FISH), Horseradish Peroxidase (HRP) and Poly-HRP, Immunohistochemistry (IHC), Physiological Probes, Power Styramide™ Signal Amplification (PSA™)
Molecular weight 1046.41 | Correction Factor (260 nm) 0.23 | Correction Factor (280 nm) 0.14 | Extinction coefficient (cm -1 M -1) 1000001 | Excitation (nm) 557 | Emission (nm) 570 | Quantum yield 0.641 |
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. iFluor 555 tyramide contains the bright iFluor 555 that can be readily detected with the standard Cy3 filter set. It is an excellent replacement for Alexa Fluor® 555 tyramide (Alexa Fluor® is the trade mark of ThermoFisher) or other spectrally similar fluorescent tyramide conjugates or TSA reagents (such as Cy3 tyramide).
Platform
Fluorescence microscope
Excitation | Cy3/TRITC filter set |
Emission | Cy3/TRITC filter set |
Recommended plate | Black wall/clear bottom |
Instrument specification(s) | Cy3/TRITC filter set |
Calculators
Common stock solution preparation
Table 1. Volume of DMSO needed to reconstitute specific mass of iFluor® 555 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 | 95.565 µL | 477.824 µL | 955.648 µL | 4.778 mL | 9.556 mL |
5 mM | 19.113 µL | 95.565 µL | 191.13 µL | 955.648 µL | 1.911 mL |
10 mM | 9.556 µL | 47.782 µL | 95.565 µL | 477.824 µL | 955.648 µL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Spectrum
Open in Advanced Spectrum Viewer


Spectral properties
Correction Factor (260 nm) | 0.23 |
Correction Factor (280 nm) | 0.14 |
Extinction coefficient (cm -1 M -1) | 1000001 |
Excitation (nm) | 557 |
Emission (nm) | 570 |
Quantum yield | 0.641 |
Product Family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield | Correction Factor (260 nm) | Correction Factor (280 nm) |
iFluor® 555 maleimide | 557 | 570 | 1000001 | 0.641 | 0.23 | 0.14 |
iFluor® 555 amine | 557 | 570 | 1000001 | 0.641 | 0.23 | 0.14 |
iFluor® 555 hydrazide | 557 | 570 | 1000001 | 0.641 | 0.23 | 0.14 |
iFluor® 488 tyramide | 491 | 516 | 750001 | 0.91 | 0.21 | 0.11 |
iFluor® 555 alkyne | 557 | 570 | 1000001 | 0.641 | 0.23 | 0.14 |
iFluor® 555 azide | 557 | 570 | 1000001 | 0.641 | 0.23 | 0.14 |
iFluor® 555 Styramide *Superior Replacement for Alexa Fluor 555 tyramide and Opal 570* | 557 | 570 | 1000001 | 0.641 | 0.23 | 0.14 |
iFluor® 647 Tyramide | 656 | 670 | 2500001 | 0.251 | 0.03 | 0.03 |
iFluor® 350 Tyramide | 345 | 450 | 200001 | 0.951 | 0.83 | 0.23 |
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Images

Figure 1. Formalin-fixed, paraffin-embedded (FFPE) human lung tissue was labeled with anti-EpCAM mouse mAb followed by HRP-labeled goat anti-mouse IgG (Cat No. 16728). The fluorescence signal was developed using iFluor® 555 tyramide (Cat No. 45105) and detected with a TRITC/Cy3 filter set. Nuclei (blue) were counterstained with DAPI (Cat No. 17507).

Figure 2. Superior sensitivity with iFluor® 555 tyramide. HeLa cells were incubated with primary anti-tubulin antibodies followed by detection with HRP-Goat anti-Mouse IgG and iFluor® 555 tyramide (Left) or Alexa Fluor® 555 tyramide (Right). Fluorescence images were taken on a Keyence BZ-X710 fluorescence microscope equipped with a Cy3 filter set.