5-FITC-Tetrazine

Guide (The following is our recommended solution. This solution merely provides guidance; it should be modified according to your specific needs.)
1. Reagent Preparation
1.1 Reconstitution/Preparation: Dissolve 5-FITC-Tetrazine in anhydrous DMSO or DMF to prepare a 1-10 mM stock solution.
1.2 Storage: Store the dry powder and reserve solution at -20°C in a dark place. Pay attention to avoiding light and moisture to prevent the degradation of the tetrazine and fluorescent core.
1.3 Buffer: Use PBS or compatible neutral physiological aqueous buffer (pH 7.2 - 7.4). If the previous steps involve free isothiocyanate reactive activity, avoid using buffers containing primary amines (such as Tris or glycine).
2. Labelling Procedure
2.2 Target Preparation: Ensure that your protein, antibody, or cell surface target has been labeled with a strained diene acceptor tag (such as TCO).
2.3 Incubation/Loading: Add the 5-FITC-Tetrazine stock solution to the TCO-labeled biological molecule solution (usually using a 2 to 5-fold molar excess of the dye).
2.4 Reaction conditions: Incubate at room temperature for 5 to 30 minutes (or up to 1 hour at 4°C). The IEDDA reaction can proceed rapidly without the need for any toxic metal catalyst.
2.5 Purification: Remove the unreacted free dyes through size exclusion chromatography (centrifugal column) or by thorough dialysis in PBS.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.