XFD488 tyramide reagent *Same Structure to Alexa Fluor™ 488 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 | 856.02 |
Solvent | DMSO |
Correction Factor (260 nm) | 0.3 |
Correction Factor (280 nm) | 0.11 |
Extinction coefficient (cm -1 M -1) | 73000 |
Excitation (nm) | 499 |
Emission (nm) | 520 |
Quantum yield | 0.921 |
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 856.02 | Correction Factor (260 nm) 0.3 | Correction Factor (280 nm) 0.11 | Extinction coefficient (cm -1 M -1) 73000 | Excitation (nm) 499 | Emission (nm) 520 | Quantum yield 0.921 |
Platform
Fluorescence microscope
Excitation | FITC filter set |
Emission | FITC filter set |
Recommended plate | Black wall/clear bottom |
Instrument specification(s) | FITC 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 100 µL DMSO to vial and mix well. Note: Unused Tyramide stock solution can be stored at 2-8 °C.
PREPARATION OF WORKING SOLUTION
Add 100 µL of Tyramide stock solution into 20 mL of buffer of your choice containing 0.003% H2O2. Note: Tris Buffer, pH=7.4 can be used for similar performance. Note: Tyramide working solution should be used immediately and made fresh on the day of use. Note: 20 mL solution is good for 200 tests.
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 | 116.82 µL | 584.099 µL | 1.168 mL | 5.841 mL | 11.682 mL |
5 mM | 23.364 µL | 116.82 µL | 233.639 µL | 1.168 mL | 2.336 mL |
10 mM | 11.682 µL | 58.41 µL | 116.82 µL | 584.099 µL | 1.168 mL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Spectrum

Spectral properties
Correction Factor (260 nm) | 0.3 |
Correction Factor (280 nm) | 0.11 |
Extinction coefficient (cm -1 M -1) | 73000 |
Excitation (nm) | 499 |
Emission (nm) | 520 |
Quantum yield | 0.921 |
Product Family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield | Correction Factor (260 nm) | Correction Factor (280 nm) |
XFD546 tyramide reagent *Same Structure to Alexa Fluor™ 546 tyramide* | 561 | 572 | 112000 | 0.791 | 0.21 | 0.12 |
XFD594 tyramide reagent *Same Structure to Alexa Fluor™ 594 tyramide* | 590 | 618 | 92000 | 0.661 | 0.43 | 0.56 |
XFD350 tyramide reagent *Same Structure to Alexa Fluor™ 350 tyramide* | 343 | 441 | 19000 | - | 0.25 | 0.19 |
XFD568 tyramide reagent *Same Structure to Alexa Fluor™ 568 tyramide* | 579 | 603 | 88000 | 0.691 | 0.45 | 0.46 |
Citations
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: Skop, Ahna and Park, Sungjin and Dahn, Randall and Kurt, Elif and Presle, Adrien and VandenHeuvel, Kathryn and Moravec, Cara and Jambhekar, Ashwini and Olukoga, Olushola and Shepherd, Jason and others,
Journal: (2022)
Authors: Huang, Jingjun and Wei, Qianhao and Zhao, Mi and Zhou, Libin and Shi, Herong and Zhang, Yong and Chen, Huapu
Journal: Aquaculture Reports (2021): 100754
Authors: Liu, P., Li, C., Zhang, R., Tang, Q., Wei, J., Lu, Y., Shen, P.
Journal: Biosens Bioelectron (2019): 543-550
Authors: Zhou, X., Li, Y., Wu, H., Huang, W., Ju, H., Ding, S.
Journal: Biosens Bioelectron (2019): 88-94
Authors: Li, X., Chen, B., He, M., Xiao, G., Hu, B.
Journal: Talanta (2018): 40-46
Authors: Rees, J. S., Li, X. W., Perrett, S., Lilley, K. S., Jackson, A. P.
Journal: Curr Protoc Protein Sci (2017): 19 27 1-19 27 18
Authors: J, undefined and ura, A., Hu, J., Wilk, R., Krause, H. M.
Journal: J Vis Exp (2017): se name="11070.enl" path="C:\Website\Referenc
Authors: Akama, K., Shirai, K., Suzuki, S.
Journal: Anal Chem (2016): 7123-9
Authors: Rees, J. S., Li, X. W., Perrett, S., Lilley, K. S., Jackson, A. P.
Journal: Curr Protoc Protein Sci (2015): 19 27 1-18
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