The orphan G protein-coupled receptor 35 (GPR35) is a potential target for the treatment of pain, inflammation, and metabolic diseases. Although many GPR35 agonists have been discovered, research on functional GPR35 ligands, such as fluorescent probes, is still limited. Herein, we developed a series of GPR35 fluorescent probes by conjugating a BODIPY fluorophore to <b>DQDA</b>, a known GPR35 agonist. All probes exhibited excellent GPR35 agonistic activity and desired spectroscopic properties, as determined by the DMR assay, bioluminescence resonance energy transfer (BRET)-based saturation, and kinetic binding experiments. Notably, compound <b>15</b> showed the highest binding potency and the weakest nonspecific BRET binding signal (<i>K</i> <sub>d</sub> = 3.9 nM). A BRET-based competition binding assay with <b>15</b> was also established and used to determine the binding constants and kinetics of unlabeled GPR35 ligands.