iFluor® 647 Tyramide
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Catalog Number | |
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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 | 1092.20 |
Solvent | DMSO |
Correction Factor (260 nm) | 0.03 |
Correction Factor (280 nm) | 0.03 |
Correction Factor (656 nm) | 0.0793 |
Extinction coefficient (cm -1 M -1) | 2500001 |
Excitation (nm) | 656 |
Emission (nm) | 670 |
Quantum yield | 0.251 |
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 | ![]() ![]() |
Molecular weight 1092.20 | Correction Factor (260 nm) 0.03 | Correction Factor (280 nm) 0.03 | Correction Factor (656 nm) 0.0793 | Extinction coefficient (cm -1 M -1) 2500001 | Excitation (nm) 656 | Emission (nm) 670 | Quantum yield 0.251 |
Platform
Fluorescence microscope
Excitation | Cy5 filter set |
Emission | Cy5 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 as 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 preferred specific solution/protocol as needed. A protocol can be found at:
https://www.aatbio.com/resources/guides/paraffin-embedded-tissue-immunohistochemistry-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 using a 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 non-specific signal, you can shorten the incubation time with Tyramide. You should optimize the incubation period using positive and negative control samples at various incubation time points. Or you can use lower concentration of Tyramide 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 counterstaining.
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 | 91.558 µL | 457.792 µL | 915.583 µL | 4.578 mL | 9.156 mL |
5 mM | 18.312 µL | 91.558 µL | 183.117 µL | 915.583 µL | 1.831 mL |
10 mM | 9.156 µL | 45.779 µL | 91.558 µL | 457.792 µL | 915.583 µL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Spectrum

Spectral properties
Correction Factor (260 nm) | 0.03 |
Correction Factor (280 nm) | 0.03 |
Correction Factor (656 nm) | 0.0793 |
Extinction coefficient (cm -1 M -1) | 2500001 |
Excitation (nm) | 656 |
Emission (nm) | 670 |
Quantum yield | 0.251 |
Product Family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield | Correction Factor (260 nm) | Correction Factor (280 nm) |
iFluor® 647 maleimide | 656 | 670 | 2500001 | 0.251 | 0.03 | 0.03 |
iFluor® 647 amine | 656 | 670 | 2500001 | 0.251 | 0.03 | 0.03 |
iFluor® 647 hydrazide | 656 | 670 | 2500001 | 0.251 | 0.03 | 0.03 |
iFluor® 647 alkyne | 656 | 670 | 2500001 | 0.251 | 0.03 | 0.03 |
iFluor® 647 azide | 656 | 670 | 2500001 | 0.251 | 0.03 | 0.03 |
iFluor® 488 tyramide | 491 | 516 | 750001 | 0.91 | 0.21 | 0.11 |
iFluor® 647 Styramide *Superior Replacement for Alexa Fluor 647 tyramide* | 656 | 670 | 2500001 | 0.251 | 0.03 | 0.03 |
iFluor® 555 Tyramide | 557 | 570 | 1000001 | 0.641 | 0.23 | 0.14 |
iFluor® 350 Tyramide | 345 | 450 | 200001 | 0.951 | 0.83 | 0.23 |
Show More (8) |
Images


Citations
Authors: Dai, Zan and Wang, Qiaoyun and Tang, Jie and Wu, Min and Li, Haoze and Yang, Yannan and Zhen, Xu and Yu, Chengzhong
Journal: Biomaterials (2022): 121261
Authors: Mohale, Mamello and Gundampati, Ravi Kumar and Kumar, Thallapuranam Krishnaswamy Suresh and Heyes, Colin D
Journal: Analytical biochemistry (2022): 114524
Authors: Verisqa, Fiona and Cha, Jae-Ryung and Nguyen, Linh and Kim, Hae-Won and Knowles, Jonathan C
Journal: Biomolecules (2022): 1692
Authors: Ji, Tengfei and Ma, Keqiang and Wu, Hongsheng and Cao, Tiansheng
Journal: (2021)
Authors: Gerlach, Brennan D and Ampomah, Patrick B and Yurdagul Jr, Arif and Liu, Chuang and Lauring, Max C and Wang, Xiaobo and Kasikara, Canan and Kong, Na and Shi, Jinjun and Tao, Wei and others,
Journal: Cell metabolism (2021): 2445--2463
Authors: Ilnytska, Olga and Lai, Kimberly and Gorshkov, Kirill and Schultz, Mark L and Tran, Bruce Nguyen and Jeziorek, Maciej and Kunkel, Thaddeus J and Azaria, Ruth D and McLoughlin, Hayley S and Waghalter, Miriam and others,
Journal: Journal of Biological Chemistry (2021)
Authors: Masibag, Angelique N and Bergin, Christopher J and Haebe, Joshua R and Zouggar, A{\"\i}cha and Shah, Muhammad S and Sandouka, Tamara and da Silva, Amanda Mendes and Desrochers, Fran{\c{c}}ois M and Fournier-Morin, Aube and Benoit, Yannick D
Journal: Iscience (2021): 103442
Authors: Li, Dong and Zhuang, Jie and He, Haisheng and Jiang, Sifan and Banerjee, Amrita and Lu, Yi and Wu, Wei and Mitragotri, Samir and Gan, Li and Qi, Jianping
Journal: ACS applied materials \& interfaces (2017): 42492--42502
References
Authors: Huang, Xiangyi and Wang, Jinjie and Liu, Heng and Lan, Tao and Ren, Jicun
Journal: Talanta (2013): 79-84
Authors: Nakamura, Ayako and Uchihara, Toshiki
Journal: Journal of neuroscience methods (2004): 67-70
Authors: Pougnard, Claire and Catala, Philippe and Drocourt, Jean-Louis and Legastelois, Stephane and Pernin, Pierre and Pringuez, Emmanuelle and Lebaron, Philippe
Journal: Applied and environmental microbiology (2002): 3102-7
Authors: Webb, D R and Bonfiglioli, R G and Carraro, L and Osler, R and Symons, R H
Journal: Phytopathology (1999): 894-901
Application notes
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