Cy3 tyramide
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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 |
Molecular weight | 863.96 |
Solvent | DMSO |
Correction Factor (260 nm) | 0.07 |
Correction Factor (280 nm) | 0.073 |
Extinction coefficient (cm -1 M -1) | 1500001 |
Excitation (nm) | 555 |
Emission (nm) | 569 |
Quantum yield | 0.151 |
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 |
iFluor® 555 Tyramide |
Overview | ![]() ![]() |
Molecular weight 863.96 | Correction Factor (260 nm) 0.07 | Correction Factor (280 nm) 0.073 | Extinction coefficient (cm -1 M -1) 1500001 | Excitation (nm) 555 | Emission (nm) 569 | Quantum yield 0.151 |
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 |
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 appropriate amount of DMSO to make 5-10 mM of Tyramide stock solution. Note: Unused Tyramide stock solution can be stored at 2-8 °C.
PREPARATION OF WORKING SOLUTION
Add 1 µL of Tyramide stock solution into 1 mL of buffer of your choice containing 0.003% H2O2. Note: Tris Buffer, pH=7.4 can be used for optimal performance. Note: Tyramide working solution should be used immediately and made fresh on the day of use.
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.
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 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.
- 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 | 115.746 µL | 578.73 µL | 1.157 mL | 5.787 mL | 11.575 mL |
5 mM | 23.149 µL | 115.746 µL | 231.492 µL | 1.157 mL | 2.315 mL |
10 mM | 11.575 µL | 57.873 µL | 115.746 µL | 578.73 µL | 1.157 mL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Spectrum

Spectral properties
Correction Factor (260 nm) | 0.07 |
Correction Factor (280 nm) | 0.073 |
Extinction coefficient (cm -1 M -1) | 1500001 |
Excitation (nm) | 555 |
Emission (nm) | 569 |
Quantum yield | 0.151 |
Product Family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield | Correction Factor (260 nm) | Correction Factor (280 nm) |
Cy3 tetrazine | 555 | 569 | 1500001 | 0.151 | 0.07 | 0.073 |
Cy3 phosphoramidite | 555 | 569 | 1500001 | 0.151 | 0.07 | 0.073 |
Cy3 aldehyde | 555 | 569 | 1500001 | 0.151 | 0.07 | 0.073 |
DBCO-Cy3 | 555 | 569 | 1500001 | 0.151 | 0.07 | 0.073 |
Cy5 tyramide | 651 | 670 | 2500001 | 0.271, 0.42 | 0.02 | 0.03 |
Cy7 tyramide | 756 | 779 | 250000 | 0.3 | 0.05 | 0.036 |
XFD514 tyramide | 518 | 543 | 80000 | - | 0.31 | 0.18 |
XFD532 tyramide | 534 | 553 | 81000 | 0.611 | 0.24 | 0.09 |
Fluorescein Tyramide | 498 | 517 | 800001 | 0.79001, 0.952 | 0.32 | 0.35 |
Images
Citations
Authors: Tian, E and Syed, Zulfeqhar A and Edin, Matthew L and Zeldin, Darryl C and Ten Hagen, Kelly G
Journal: Glycobiology (2023): cwad031
Authors: Luo, Haiyun and Liu, Wenjing and Zhou, Yachuan and Zhang, Yanli and Wu, Junrong and Wang, Ruolan and Shao, Longquan
Journal: Journal of Translational Medicine (2022): 1--15
Authors: Mizoguchi, Takamasa and Fukada, Michi and Iihama, Miku and Song, Xuehui and Fukagawa, Shun and Kuwabara, Shuhei and Omaru, Shuhei and Higashijima, Shin-ichi and Itoh, Motoyuki
Journal: Development (2020): dev191312
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