Tyramide (fluorescently labeled)

MATERIALS

METHOD

Perform procedure in the hood. For best results, use fresh reagents. NHS esters are unstable, and the coupling reaction should be kept anhydrous.

• 1. Dissolve 40 mg of fluorescently labeled NHS ester in 4 mL of DMF.

• 2. Add 10 μL of TEN to 1 mL of DMF.

• 3. Dissolve 10 mg of tyramide in 1 mL of TEN-DMF solution.

• 4. Mix 4 mL of fluorescently labeled NHS ester in DMF from Step 1 with 1.37 mL of tyramide solution from Step 3. Incubate in the dark for 2 h at room temperature.

• 5. Add 4.6 mL of ethanol.

• 6. Keep protected from light. Store at 4°C or -20°C.

  The solution is stable for at least 8 mo at 4°C.