CY3-YNE (solution)

Guidelines (The following is our recommended protocol, which serves only as a guideline and should be modified according to your specific needs)^[1].
Specific steps:
1) Premix CuSO₄ (20 mM aqueous solution) and BTTP (20 mM DMSO solution) at a ratio of 1:4 (v/v) to form the Cu (II)-BTTP complex.
2) Add 2 μL of Cu (II)-BTTP to a solution containing 83 μL of PBS buffer, 4 μL of DMSO and 1 μL of CY3-YNE (10 mM DMSO solution). Then, add 10 μL of freshly prepared sodium ascorbate (25 mM aqueous solution) and mix thoroughly (final concentrations: 100 μM CY3-YNE, 160 μM CuSO₄, 640 μM BTTP, 2.5 mM sodium ascorbate). A separate solution without Cu (II)-BTTP is used as the control. Inject the solutions into the channels and incubate for 1 h at room temperature in the dark.
3) After the reaction, thoroughly wash the channels with water and blow dry with nitrogen.
4) Acquire fluorescence images of the chip using a Sapphire biomolecular imager.
5) The imager settings are as follows: Dye: Cy3, Intensity: 5, Pixel size: 500 μm, Scan speed: High, Sample type: Slide.
6) To quantify the areal density of surface functional groups, scan a series of dilutions of CY3-YNE using the same settings.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.