BODIPY FL prazosin is a fluorescent α1-adrenergic antagonist with binding affinities of Ki: 14.5 nM for α1a-AR and Ki: 43.3 nM for α1b-AR, used to study subcellular localization differences in α1-adrenoceptor subtypes.

α1B-adrenergic receptor:43.3 nM (Ki), α1A-adrenergic receptor:14.5 nM (Ki)

BODIPY FL prazosin (10 nM; 30 min at room temperature in 100 μl; COS-7 cells) shows Affinity of various α1-AR ligands with K i values of 14.5, 43.3 nM for α1a-AR and α1b-AR, respectively [1]. BODIPY FL prazosin (100 nM, 30 min) can be used as molecular probe for the Visualization of the non-adrenoceptor binding site of α1-adrenergic drugs in erythroleukemia cells [3].

Instructions for use
I. Solution preparation
1. Mother solution dissolution and preparation: BODIPY FL prazosin is recommended to be dissolved in anhydrous DMSO. The concentration can be adjusted according to the experimental requirements, usually between 10-50 μM.
2. Preparation of working solution: It can be diluted to the required concentration with buffer or culture medium according to the experimental needs. Usually, a certain preliminary experiment can be performed before use to optimize the final concentration.
II. Operation steps
1. Labeling reaction:
1) Add BODIPY FL prazosin solution (1-10uM) to the cell culture system. Generally, it is cultured at 37°C for a certain period of time (30 minutes to 1 hour) to ensure that the fluorescent label binds to the α1-AR on the cell surface.
2) During the labeling process, it should be ensured that the labeling agent does not have a toxic effect on the cells. It is recommended to use appropriate control experiments.
2. Cleaning and analysis:
1) After labeling, use an appropriate washing buffer to remove unbound fluorescent ligands to reduce background signals.
2) When performing flow cytometry or confocal microscopy analysis, ensure that the excitation light source and filter settings are correct to accurately collect fluorescence signals in the 485/535 nm wavelength range.
3. Analysis:
1) Use flow cytometry for quantitative analysis and evaluate the expression level of α1-AR by intracellular fluorescence intensity.
2) When using confocal microscopy, fluorescently labeled cells can be observed at high resolution to study the distribution and localization of receptor subtypes.
Notes:
1. BODIPY FL prazosin should be stored at -20°C to avoid repeated freezing and thawing.
2. The solution used in the experiment should be as fresh as possible and avoid long-term exposure to strong light to maintain its fluorescence stability.