iFluor® 546 Tyramide
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Catalog Number | |
Unit Size | |
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Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Molecular weight | 1282.88 |
Solvent | DMSO |
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 |
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 | ![]() ![]() |
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 |
Platform
Fluorescence microscope
Excitation | Cy3/TRITC filter set |
Emission | Cy3/TRITC filter set |
Recommended plate | Black wall/clear bottom |
Example protocol
AT A GLANCE
- Fix/permeabilize/block cells or tissue
- Add primary antibody in blocking buffer
- Add HRP-conjugated secondary antibody
- Prepare tyramide working solution and apply in cells or tissue for 5-10 minutes at room temperature
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
Add 100 µL of DMSO to the vial of iFluor® tyramide and mix well.
Note: Make single-use aliquots and store unused 200X stock solution at 2-8 °C, protected from light. Avoid repeat freeze-thaw cycles.
PREPARATION OF WORKING SOLUTION
Add 100 µL of the tyramide stock solution into 20 mL of a buffer of your choice containing 0.003% H2O2.
Note: For optimal performance, use Tris Buffer, pH=7.4.
Note: A 20 mL solution is good for 200 tests. The tyramide working solution should be used immediately and made fresh on the day of use. Avoid direct exposure to light.
Make an appropriate concentration of secondary antibody-HRP working solution per the manufacturer's recommendations.
SAMPLE EXPERIMENTAL PROTOCOL
This protocol is applicable for both cells and tissues staining.
- Fix the cells or tissue with 3.7% formaldehyde or paraformaldehyde, in PBS at room temperature for 20 minutes.
- Rinse the cells or tissue with PBS twice.
- Permeabilize the cells with 0.1% Triton X-100 solution for 1-5 minutes at room temperature.
- Rinse the cells or tissue with PBS twice.
Deparaffinize and dehydrate the tissue according to the standard IHC protocols. Perform antigen retrieval with the preferred specific solution/protocol as needed. A protocol can be found at:
https://www.aatbio.com/resources/guides/paraffin-embedded-tissueimmunohistochemistry-protocol.html
- Optional: Quench endogenous peroxidase activity by incubating cell or tissue sample in peroxidase quenching solution (such as 3% hydrogen peroxide) for 10 minutes. Rinse with PBS twice at room temperature.
- Optional: If using HRP-conjugated streptavidin, it is advisable to block endogenous biotins by biotin blocking buffer.
- Block with preferred blocking solution (such as PBS with 1% BSA) for 30 minutes at 4 °C.
- Remove blocking solution and add primary antibody diluted in recommended antibody diluent for 60 minutes at room temperature or overnight at 4 °C.
- Wash with PBS three times for 5 minutes each.
Apply 100 µL of secondary antibody-HRP working solution to each sample and incubate for 60 minutes at room temperature.
Note: Incubation time and concentration can be varied depending on the signal intensity.
- Wash with PBS three times for 5 minutes each.
Prepare and apply 100 µL of Tyramide working solution to each sample and incubate for 5-10 minutes at room temperature.
Note: If you observe a non-specific signal, you can shorten the incubation time with the tyramide reagent. You should optimize the incubation period using positive and negative control samples at various incubation time points. Or you can use a lower concentration of the tyramide reagent in the working solution.
- Rinse with PBS three times.
- Counterstain the cell or tissue samples as needed. AAT provides a series of nucleus counterstain reagents as listed in Table 1. Follow the instruction provided with the reagents.
Mount the coverslip using a mounting medium with anti-fading properties.
Note: To ensure optimal results, it is recommended to use either ReadiUse™ microscope mounting solution (Cat. 20009) or FluoroQuest™ TSA/PSA Antifade Mounting Medium *Optimized for Tyramide and Styramide Imaging* (Cat. 44890) instead of Vectashield® mounting media. There are instances where Vectashield® mounting media may not be suitable for certain TSA/PSA conjugates.
- Use the appropriate filter set to visualize the signal from the Tyramide labeling.
Table 1. Products recommended for nucleus counterstain
Cat# | Product Name | Ex/Em (nm) |
17548 | Nuclear Blue™ DCS1 | 350/461 |
17550 | Nuclear Green™ DCS1 | 503/526 |
17551 | Nuclear Orange™ DCS1 | 528/576 |
17552 | Nuclear Red™ DCS1 | 642/660 |
Calculators
Common stock solution preparation
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
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
/ | = | x | = |
Spectrum

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


References
Authors: Lyu, Qiongxia and Zheng, Huifei Sophia and Laprocina, Karly and Huang, Chen-Che Jeff
Journal: Journal of visualized experiments : JoVE (2020)
Authors: Wang, Herui and Pangilinan, Ryan L and Zhu, Yan
Journal: Methods in molecular biology (Clifton, N.J.) (2020): 89-97
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: Lauter, Gilbert and Söll, Iris and Hauptmann, Giselbert
Journal: Methods in molecular biology (Clifton, N.J.) (2020): 397-409
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: 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: 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: 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: Khrustaleva, Ludmila and Kudryavtseva, Natalia and Romanov, Dmitry and Ermolaev, Aleksey and Kirov, Ilya
Journal: Scientific reports (2019): 12007
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
Application notes
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FAQ
Are there any alternatives to BrdU (Bromodeoxyuridine)?
Are there any alternatives to Cy5?
Are there any alternatives to indocyanine green (ICG)?
Are there any calcium indicators that don't require probenecid (PBC)?