650 compound 1 nhs

EXAMPLE 30

The inventive compounds and commercial dye were evaluated in direct fluorescence labeling of cell surface proteins. DyLight 649-NHS, 650 Compound 1-NHS, and Whole Cell Stain Orange (Thermo Fisher Scientific) were reconstituted in DMF and diluted to 6 μM in Dulbecco's PBS (DPBS). A total of four 1:1 serial dilutions of the dyes were prepared in DPBS. Frozen IMR90 cells (human lung embryonic fibroblast) on a plate were thawed for 1 h at 37° C. The cell plates were washed two times with DPBS and incubated with diluted dye for 30 min at room temperature, protected from light. The cell plates were then washed three times with DPBS. The cell plates were incubated with 100 μl/well of 1 μg/ml Hoechst dye in DPBS. The cell plates were sealed and imaged using the Thermo Scientific ArrayScan VTI HCS Reader.

As shown in FIG. 51, 650 Compound 1-NHS (row 3) performed equivalently as DyLight 649-NHS (row 2) and Whole Cell Stain Orange (row 1) at 6 μm (column A), 3 μm (column B), 1.5 μm (column C), and 0.75 μm (column D).

EXAMPLE 30

The inventive compounds and commercial dye were evaluated in direct fluorescence labeling of cell surface proteins. DyLight 649-NHS, 650 Compound 1-NHS, and Whole Cell Stain Orange (Thermo Fisher Scientific) were reconstituted in DMF and diluted to 6 μM in Dulbecco's PBS (DPBS). A total of four 1:1 serial dilutions of the dyes were prepared in DPBS. Frozen IMR90 cells (human lung embryonic fibroblast) on a plate were thawed for 1 h at 37° C. The cell plates were washed two times with DPBS and incubated with diluted dye for 30 min at room temperature, protected from light. The cell plates were then washed three times with DPBS. The cell plates were incubated with 100 μl/well of 1 μg/ml Hoechst dye in DPBS. The cell plates were sealed and imaged using the Thermo Scientific ArrayScan VTI HCS Reader.

As shown in FIG. 51, 650 Compound 1-NHS (row 3) performed equivalently as DyLight 649-NHS (row 2) and Whole Cell Stain Orange (row 1) at 6 μm (column A), 3 μm (column B), 1.5 μm (column C), and 0.75 μm (column D).

EXAMPLE 30

The inventive compounds and commercial dye were evaluated in direct fluorescence labeling of cell surface proteins. DyLight 649-NHS, 650 Compound 1-NHS, and Whole Cell Stain Orange (Thermo Fisher Scientific) were reconstituted in DMF and diluted to 6 μM in Dulbecco's PBS (DPBS). A total of four 1:1 serial dilutions of the dyes were prepared in DPBS. Frozen IMR90 cells (human lung embryonic fibroblast) on a plate were thawed for 1 h at 37° C. The cell plates were washed two times with DPBS and incubated with diluted dye for 30 min at room temperature, protected from light. The cell plates were then washed three times with DPBS. The cell plates were incubated with 100 μl/well of 1 μg/ml Hoechst dye in DPBS. The cell plates were sealed and imaged using the Thermo Scientific ArrayScan VTI HCS Reader.

As shown in FIG. 51, 650 Compound 1-NHS (row 3) performed equivalently as DyLight 649-NHS (row 2) and Whole Cell Stain Orange (row 1) at 6 μm (column A), 3 μm (column B), 1.5 μm (column C), and 0.75 μm (column D).