TP-α

Guidelines (The following are our recommended protocols; this is a guideline and should be modified according to your specific needs).
1. Stock Solution Preparation
1.1 Solvent: For most dyes, organic solvents such as anhydrous DMSO are typically used for dissolution.
1.2 Concentration Recommendation: Generally, a high concentration of 1-10 mM stock solution is recommended.
1.3 Storage Conditions: Store at -20 ℃ or -80 ℃ protected from light, and repeated freeze-thaw cycles should be avoided.
2. Working Solution Preparation
2.1 Diluent: Serum-free medium or PBS is typically used. Proteins and esterases in serum may interfere with staining or cause dye hydrolysis.
2.2 Working Concentration: 0.5-10 μM
Note: Please adjust the dye working solution concentration according to your actual situation, and prepare fresh before use.
3. Cell Staining
3.1 Culture cells on sterile coverslips.
3.2 Remove the coverslips from the culture medium and aspirate excess medium.
3.3 Add working solution, gently shake to completely cover the cells, and incubate at room temperature for 1 h.
3.4 Aspirate the dye working solution, wash 2-3 times with culture medium, 5 minutes each time, and observe using a fluorescence microscope or flow cytometry.
Note: If flow cytometry is required, the cells should be digested and resuspended with trypsin before staining.

TP-α (1-2 μM; 1-24 h) has minimal cytotoxicity toward alpha TC1 and beta TC6 cells after incubation for 1 or 24 h^[1].

TP-α (0.5-1 μM; 1 h) selectively stains glucagon-secreting alpha TC1 cells, with negligible fluorescence in beta TC6 and acinar 266-6 cells^[1].
TP-α (10 μM; 30 min on ice) binds directly and selectively to glucagon with a K_d of 65 μM, showing a 9-fold fluorescence increase at 1 mg/mL glucagon^[1].