AF405 DBCO

Guidelines (The recommended experimental protocol below is for guidance only and should be modified according to your specific requirements)
1. General Reaction Conditions
1.1 Reaction Type: SPAAC (Copper-Free Click Chemistry)
1.2 Buffer: Phosphate-buffered saline (PBS) or other amine-free, neutral pH buffers (pH 7.2-7.4). Avoid using primary or secondary amine additives in storage buffers.
1.3 Temperature: Room temperature (19°C-25°C), or 4°C for sensitive targets.
1.4 Reaction Time: 2 hours to overnight, depending on the steric accessibility of the azide group.

2. Step-by-Step Labeling Protocol
2.1 Prepare Azide Target: Ensure that biomolecules (proteins, antibodies or cell surface glycans) carry reactive azide groups (-N₃) in an optimized buffer.
2.2 Reconstitute Dye: Dissolve AF405 DBCO in anhydrous DMSO or DMF (HY-Y0345) to prepare a stock solution with a concentration of 1-10 mM.
2.3 Mix Components: Add AF405 DBCO at a 2- to 10-fold molar excess to the azide-labeled biomolecule solution.
2.4 Incubation: Protect the reaction tube from light and incubate at room temperature for 2 hours (or at 4°C overnight).
2.5 Purification: Remove unreacted free dye using spin desalting columns, dialysis or size-exclusion chromatography.