| Bioactivity | Cy3 methyltetrazine (TZ-Cy3) is a click chemistry reagent with methyltetrazine building blocks that is highly reactive towards cyclooctene. Cy3 methyltetrazine is also a tetrazine-modified fluorescent probe that can be used to analyze protein phosphorylation in solution and living cells[1]. |
| Invitro | Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).Confocal imaging of phosphorylation in cells:1. Culture H1299 cells in a coverglass-bottom imaging dish for 24 h (take H1299 cells as an example).2. Incubate H1299 cells with different concentrations of ATP-NB (a ATP analogue functionalized by norbornene, which can penetrate cells and efficiently phosphorylate proteins in living cells) for 1 h.3. Add Cy3 methyltetrazine to label the phosphorylated protein with fluorescence.4. Incubate with fresh medium for 1 h.5. Wash the cells twice with PBS to release the free Cy3 methyltetrazine probe.6.Incubate the cells with 4% paraformaldehyde for 15 min and with 0.1% Triton X-100 solution at room temperature for 10 min.7. Wash cells 4 times with PBS.8. Use a confocal microscopy imaging system to obtain the high-resolution images of cells (Ex=488 nm; Em=561 nm). |
| Name | Cy3 methyltetrazine |
| CAS | 2183473-57-4 |
| Formula | C40H46BF4N7O |
| Molar Mass | 727.64 |
| Transport | Room temperature in continental US; may vary elsewhere. |
| Storage | Please store the product under the recommended conditions in the Certificate of Analysis. |