iFluor® 546 Tyramide
Ordering information
Price | |
Catalog Number | |
Unit Size | |
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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 | 1282.88 |
Solvent | DMSO |
Spectral properties
Correction Factor (260 nm) | 0.25 |
Correction Factor (280 nm) | 0.15 |
Extinction coefficient (cm -1 M -1) | 1000001 |
Excitation (nm) | 541 |
Emission (nm) | 557 |
Quantum yield | 0.671 |
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 | 12171501 |
Overview | ![]() ![]() |
See also: Antibodies and Proteomics, Antibody and Protein Labeling, Bioconjugation, Horseradish Peroxidase (HRP) and Poly-HRP, Immunohistochemistry (IHC), Power Styramide™ Signal Amplification (PSA™)
Molecular weight 1282.88 | Correction Factor (260 nm) 0.25 | Correction Factor (280 nm) 0.15 | Extinction coefficient (cm -1 M -1) 1000001 | Excitation (nm) 541 | Emission (nm) 557 | Quantum yield 0.671 |
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 546 tyramide contains the bright iFluor 546 that can be readily detected with the standard TRITC filter set. It is an excellent replacement for Alexa Fluor® 546 tyramide (Alexa Fluor® is the trade mark of ThermoFisher) or other spectrally similar fluorescent tyramide conjugates or TSA reagents (such as fluorescein tyramide).
Calculators
Common stock solution preparation
Table 1. Volume of DMSO needed to reconstitute specific mass of iFluor® 546 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 | 77.95 µL | 389.748 µL | 779.496 µL | 3.897 mL | 7.795 mL |
5 mM | 15.59 µL | 77.95 µL | 155.899 µL | 779.496 µL | 1.559 mL |
10 mM | 7.795 µL | 38.975 µL | 77.95 µL | 389.748 µL | 779.496 µL |
Molarity calculator
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Spectrum
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Spectral properties
Correction Factor (260 nm) | 0.25 |
Correction Factor (280 nm) | 0.15 |
Extinction coefficient (cm -1 M -1) | 1000001 |
Excitation (nm) | 541 |
Emission (nm) | 557 |
Quantum yield | 0.671 |
Product Family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield | Correction Factor (260 nm) | Correction Factor (280 nm) |
iFluor® 488 tyramide | 491 | 516 | 750001 | 0.91 | 0.21 | 0.11 |
iFluor® 546 Styramide *Superior Replacement for Alexa Fluor 546 tyramide* | 541 | 557 | 1000001 | 0.671 | 0.25 | 0.15 |
iFluor® 555 Tyramide | 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 |
iFluor® 568 Tyramide | 568 | 587 | 1000001 | 0.571 | 0.34 | 0.15 |
iFluor® 594 Tyramide | 588 | 604 | 1800001 | 0.531 | 0.05 | 0.04 |
iFluor® 633 tyramide | 640 | 654 | 2500001 | 0.291 | 0.062 | 0.044 |
iFluor® 430 Tyramide *Superior Replacement for Opal 480* | 433 | 498 | 400001 | 0.781 | 0.68 | 0.3 |
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Images

Figure 1. Microtubules of fixed HeLa cells were labeled with anti-α tubulin mouse mAb followed by HRP-labeled goat anti-mouse IgG (Cat No. 16728). The fluorescence signal was developed using Alexa Fluor® 546 tyramide or iFluor® 546 tyramide (Cat No. 45103) and detected with a TRITC/Cy3 filter set. iFluor® 546 tyramide shows significantly higher fluorescence intensity than Alexa Fluor® 546 tyramide under the same conditions.

Figure 2. 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® 546 tyramide (Cat No. 45103) or Alexa Fluor® 546 tyramide and detected with a TRITC/Cy3 filter set. Nuclei (blue) were counterstained with DAPI (Cat No. 17507).
References
View all 50 references: Citation Explorer
Microwaving and Fluorophore-Tyramide for Multiplex Immunostaining on Mouse Adrenals - Using Unconjugated Primary Antibodies from the Same Host Species.
Authors: Lyu, Qiongxia and Zheng, Huifei Sophia and Laprocina, Karly and Huang, Chen-Che Jeff
Journal: Journal of visualized experiments : JoVE (2020)
Authors: Lyu, Qiongxia and Zheng, Huifei Sophia and Laprocina, Karly and Huang, Chen-Che Jeff
Journal: Journal of visualized experiments : JoVE (2020)
Detection of Cytokine Receptors Using Tyramide Signal Amplification for Immunofluorescence.
Authors: Wang, Herui and Pangilinan, Ryan L and Zhu, Yan
Journal: Methods in molecular biology (Clifton, N.J.) (2020): 89-97
Authors: Wang, Herui and Pangilinan, Ryan L and Zhu, Yan
Journal: Methods in molecular biology (Clifton, N.J.) (2020): 89-97
Procedural Requirements and Recommendations for Multiplex Immunofluorescence Tyramide Signal Amplification Assays to Support Translational Oncology Studies.
Authors: Parra, Edwin Roger and Jiang, Mei and Solis, Luisa and Mino, Barbara and Laberiano, Caddie and Hernandez, Sharia and Gite, Swati and Verma, Anuj and Tetzlaff, Michael and Haymaker, Cara and Tamegnon, Auriole and Rodriguez-Canales, Jaime and Hoyd, Clifford and Bernachez, Chantale and Wistuba, Ignacio
Journal: Cancers (2020)
Authors: Parra, Edwin Roger and Jiang, Mei and Solis, Luisa and Mino, Barbara and Laberiano, Caddie and Hernandez, Sharia and Gite, Swati and Verma, Anuj and Tetzlaff, Michael and Haymaker, Cara and Tamegnon, Auriole and Rodriguez-Canales, Jaime and Hoyd, Clifford and Bernachez, Chantale and Wistuba, Ignacio
Journal: Cancers (2020)
Sensitive Multiplexed Fluorescent In Situ Hybridization Using Enhanced Tyramide Signal Amplification and Its Combination with Immunofluorescent Protein Visualization in Zebrafish.
Authors: Lauter, Gilbert and Söll, Iris and Hauptmann, Giselbert
Journal: Methods in molecular biology (Clifton, N.J.) (2020): 397-409
Authors: Lauter, Gilbert and Söll, Iris and Hauptmann, Giselbert
Journal: Methods in molecular biology (Clifton, N.J.) (2020): 397-409
Optimization of prostate cancer cell detection using multiplex tyramide signal amplification.
Authors: Roy, Sounak and Axelrod, Haley D and Valkenburg, Kenneth C and Amend, Sarah and Pienta, Kenneth J
Journal: Journal of cellular biochemistry (2019): 4804-4812
Authors: Roy, Sounak and Axelrod, Haley D and Valkenburg, Kenneth C and Amend, Sarah and Pienta, Kenneth J
Journal: Journal of cellular biochemistry (2019): 4804-4812
Highly Sensitive Detection of PCV2 Based on Tyramide Signals and GNPL Amplification.
Authors: Zhang, Shouping and Hu, Bin and Xia, Xiaojing and Xu, Yanzhao and Hang, Bolin and Jiang, Jinqing and Hu, Jianhe
Journal: Molecules (Basel, Switzerland) (2019)
Authors: Zhang, Shouping and Hu, Bin and Xia, Xiaojing and Xu, Yanzhao and Hang, Bolin and Jiang, Jinqing and Hu, Jianhe
Journal: Molecules (Basel, Switzerland) (2019)
DNAzyme Catalyzed Tyramide Depositing Reaction for In Situ Imaging of Protein Status on the Cell Surface.
Authors: Xu, Lulu and Liu, Shengchun and Yang, Tiantian and Shen, Yifan and Zhang, Yuhong and Huang, Lizhen and Zhang, Lutan and Ding, Shijia and Song, Fangzhou and Cheng, Wei
Journal: Theranostics (2019): 1993-2002
Authors: Xu, Lulu and Liu, Shengchun and Yang, Tiantian and Shen, Yifan and Zhang, Yuhong and Huang, Lizhen and Zhang, Lutan and Ding, Shijia and Song, Fangzhou and Cheng, Wei
Journal: Theranostics (2019): 1993-2002
Cancer immunophenotyping by seven-colour multispectral imaging without tyramide signal amplification.
Authors: Ijsselsteijn, Marieke E and Brouwer, Thomas P and Abdulrahman, Ziena and Reidy, Eileen and Ramalheiro, Ana and Heeren, A Marijne and Vahrmeijer, Alexander and Jordanova, Ekaterina S and de Miranda, Noel Fcc
Journal: The journal of pathology. Clinical research (2019): 3-11
Authors: Ijsselsteijn, Marieke E and Brouwer, Thomas P and Abdulrahman, Ziena and Reidy, Eileen and Ramalheiro, Ana and Heeren, A Marijne and Vahrmeijer, Alexander and Jordanova, Ekaterina S and de Miranda, Noel Fcc
Journal: The journal of pathology. Clinical research (2019): 3-11
Comparative Tyramide-FISH mapping of the genes controlling flavor and bulb color in Allium species revealed an altered gene order.
Authors: Khrustaleva, Ludmila and Kudryavtseva, Natalia and Romanov, Dmitry and Ermolaev, Aleksey and Kirov, Ilya
Journal: Scientific reports (2019): 12007
Authors: Khrustaleva, Ludmila and Kudryavtseva, Natalia and Romanov, Dmitry and Ermolaev, Aleksey and Kirov, Ilya
Journal: Scientific reports (2019): 12007
An ultrasensitive electrochemical immunosensor for procalcitonin detection based on the gold nanoparticles-enhanced tyramide signal amplification strategy.
Authors: Liu, Pei and Li, Chao and Zhang, Ruixuan and Tang, Qing and Wei, Jia and Lu, Yan and Shen, Pingping
Journal: Biosensors & bioelectronics (2019): 543-550
Authors: Liu, Pei and Li, Chao and Zhang, Ruixuan and Tang, Qing and Wei, Jia and Lu, Yan and Shen, Pingping
Journal: Biosensors & bioelectronics (2019): 543-550