CX-P

Guide
(The following is our recommended protocol. This protocol is for guidance only, should be modified according to your specific needs).

1. Stock Solution Preparation
1.2 Concentration recommendation: 1 mM.

2. Working Solution Preparation
2.1 Diluent: PBS buffer solution.
2.2 Working concentration: 10 μM.
2.3 Note: Adjust working solution concentration as needed; prepare fresh before use.

3. Staining Procedure
3.1 Sample type: HeLa, 4T1, 293T cells^[1].
3.2 Incubation conditions: Incubate with 10 μM CX-P for 30 min at 37°C.
3.3 Washing steps: Wash with PBS buffer solution (pH 7.4) three times.

4. Controls
4.2 Set up negative, positive, and blank controls.
4.3 Positive control validates experimental system; blank control excludes reagent fluorescence interference.
4.3.1 Incubate cells with CX-OH (10 μM) for 30 min at 37°C as a non-targeted control.
4.3.2 Pre-treat cancer cells with 1 mM Biotin (HY-B0511) for 30 min before CX-P incubation as a blocking control.

5. Detection & Analysis
5.1 Instrument:
5.1.1 Confocal fluorescence microscopy for cell imaging: Ex = 560 nm, Em = 650-750 nm.
5.2.1 Fluorescence intensity changes: Strong red fluorescence in low-polarity environments; fluorescence intensity decreases with increasing polarity.
5.2.1.1 HeLa and 4T1 cancer cells show strong red fluorescence, while 293T normal cells show negligible fluorescence.
5.2.1.2 LPS-pretreated cancer cells show enhanced fluorescence; sucrose or taxol-pretreated cancer cells show reduced fluorescence.

CX‑P (10 μM) is barely interfered by viscosity, pH (4.0‑10.0) and multiple biologically relevant interfering small molecules^[1].