Cy7 tyramide
Ordering information
| Price | |
| Catalog Number | |
| Unit Size | |
| Quantity |
Additional ordering information
| 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 |
Physical properties
| Molecular weight | 903.21 |
| Solvent | DMSO |
Spectral properties
| Correction Factor (260 nm) | 0.05 |
| Correction Factor (280 nm) | 0.036 |
| Correction Factor (482 nm) | 0.0005 |
| Correction Factor (565 nm) | 0.0193 |
| Correction Factor (650 nm) | 0.165 |
| Extinction coefficient (cm -1 M -1) | 250000 |
| Excitation (nm) | 756 |
| Emission (nm) | 779 |
| Quantum yield | 0.3 |
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 |  SDSProtocol |
See also: Cyanines
Molecular weight 903.21 | Correction Factor (260 nm) 0.05 | Correction Factor (280 nm) 0.036 | Correction Factor (482 nm) 0.0005 | Correction Factor (565 nm) 0.0193 | Correction Factor (650 nm) 0.165 | Extinction coefficient (cm -1 M -1) 250000 | Excitation (nm) 756 | Emission (nm) 779 | Quantum yield 0.3 |
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. 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. Cy7 tyramide contains the popular Cy7 fluorophore that can be readily detected with the standard Cy7 filter set. Its NIR excitation and emission make the probe an ideal choice for the applications where the common tyramides may have an interference resulted from the inherent fluorescence of tissues or other samples.
Platform
Fluorescence microscope
| Excitation | Cy7 filter set |
| Emission | Cy7 filter set |
| Recommended plate | Black wall/clear bottom |
Example protocol
AT A GLANCE
Protocol Summary
- 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.
Tyramide stock solution (200X)
Add appropriate amount of DMSO to make 1 or 2 mg/mL stock solution and mix well. Note: Unused Tyramide stock solution can be stored at 2-8 °C.PREPARATION OF WORKING SOLUTION
Tyramide working solution (1X)
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: Concentration should be optimized initially for final appropriate concentration for the tests.SAMPLE EXPERIMENTAL PROTOCOL
This protocol is applicable for both cells and tissues staining.
Protocol can be found at: https://www.aatbio.com/resources/guides/paraffin-embedded-tissueimmunohistochemistry-protocol.html
Cell fixation and permeabilization
- 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.
Tissue fixation, deparaffinization and rehydration
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
Peroxidase labeling
- 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.
Tyramide labeling
- 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 and fluorescence imaging
- 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.
| 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
Table 1. Volume of DMSO needed to reconstitute specific mass of Cy7 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 | 110.716 µL | 553.581 µL | 1.107 mL | 5.536 mL | 11.072 mL |
| 5 mM | 22.143 µL | 110.716 µL | 221.432 µL | 1.107 mL | 2.214 mL |
| 10 mM | 11.072 µL | 55.358 µL | 110.716 µL | 553.581 µL | 1.107 mL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
| Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
| / | = | x | = |
Spectrum
Open in Advanced Spectrum Viewer
Spectral properties
| Correction Factor (260 nm) | 0.05 |
| Correction Factor (280 nm) | 0.036 |
| Correction Factor (482 nm) | 0.0005 |
| Correction Factor (565 nm) | 0.0193 |
| Correction Factor (650 nm) | 0.165 |
| Extinction coefficient (cm -1 M -1) | 250000 |
| Excitation (nm) | 756 |
| Emission (nm) | 779 |
| Quantum yield | 0.3 |
Product Family
| Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield | Correction Factor (260 nm) | Correction Factor (280 nm) |
| Cy7 Styramide *Superior Replacement for Cy7 tyramide* | 756 | 779 | 250000 | 0.3 | 0.05 | 0.036 |
| Cy3 tyramide | 555 | 569 | 1500001 | 0.151 | 0.07 | 0.073 |
| Cy5 tyramide | 651 | 670 | 2500001 | 0.271, 0.42 | 0.02 | 0.03 |
| 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 |
| PerCP-Cy7 | 482 | 784 | 350000 | - | - | - |
Images
Figure 1. Chemical structure for Cy7 tyramide.
References
View all 50 references: Citation Explorer
Tyramide Signal-Amplified Immunofluorescence of MYCN and MYC in Human Tissue Specimens and Cell Line Cultures.
Authors: Schafer, Johanna M and Pietenpol, Jennifer A
Journal: Bio-protocol (2020): e3677
Authors: Schafer, Johanna M and Pietenpol, Jennifer A
Journal: Bio-protocol (2020): e3677
Cascade signal amplification for sensitive detection of exosomes by integrating tyramide and surface-initiated enzymatic polymerization.
Authors: Huang, Zhipeng and Lin, Qiuyuan and Yang, Bin and Ye, Xin and Chen, Hui and Weng, Wenhao and Kong, Jilie
Journal: Chemical communications (Cambridge, England) (2020): 12793-12796
Authors: Huang, Zhipeng and Lin, Qiuyuan and Yang, Bin and Ye, Xin and Chen, Hui and Weng, Wenhao and Kong, Jilie
Journal: Chemical communications (Cambridge, England) (2020): 12793-12796
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
Characterizing the Tumor Immune Microenvironment with Tyramide-Based Multiplex Immunofluorescence.
Authors: Mori, Hidetoshi and Bolen, Jennifer and Schuetter, Louis and Massion, Pierre and Hoyt, Clifford C and VandenBerg, Scott and Esserman, Laura and Borowsky, Alexander D and Campbell, Michael J
Journal: Journal of mammary gland biology and neoplasia (2020): 417-432
Authors: Mori, Hidetoshi and Bolen, Jennifer and Schuetter, Louis and Massion, Pierre and Hoyt, Clifford C and VandenBerg, Scott and Esserman, Laura and Borowsky, Alexander D and Campbell, Michael J
Journal: Journal of mammary gland biology and neoplasia (2020): 417-432
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
Tyramide signal amplification mass spectrometry (TSA-MS) ratio identifies nuclear speckle proteins.
Authors: Dopie, Joseph and Sweredoski, Michael J and Moradian, Annie and Belmont, Andrew S
Journal: The Journal of cell biology (2020)
Authors: Dopie, Joseph and Sweredoski, Michael J and Moradian, Annie and Belmont, Andrew S
Journal: The Journal of cell biology (2020)
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)
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
A amperometric immunosensor for sensitive detection of circulating tumor cells using a tyramide signal amplification-based signal enhancement system.
Authors: Zhou, Xiaoyan and Li, Yujian and Wu, Haiping and Huang, Wei and Ju, Huangxian and Ding, Shijia
Journal: Biosensors & bioelectronics (2019): 88-94
Authors: Zhou, Xiaoyan and Li, Yujian and Wu, Haiping and Huang, Wei and Ju, Huangxian and Ding, Shijia
Journal: Biosensors & bioelectronics (2019): 88-94