α-Synuclein aggregate binder 1

Optical imaging of mouse brain:
α-Synuclein aggregate binder 1 (C05-05) (30μM; 30 min) specifically binds to the phosphorylated α-synuclein lesion area in the brain slices of mice inoculated with α-synuclein fibrils, and the fluorescence signal overlaps with the anti-phosphorylated α-synuclein antibody staining area^[1].
PET imaging experiment:
¹⁸F-labeled C05-05 showed appropriate brain penetration and clearance rate in normal mouse brains, and specifically aggregated in the striatum of α-synuclein fibril-inoculated mouse model^[1].
α-synuclein aggregation inhibition experiment: C05-05 (20 μM/200 μM; 24 h shaking culture) inhibited the formation of α-synuclein aggregates in a dose-dependent manner, with an inhibition rate of 60% at 200 μM^[1].

Operational steps and methods for different experimental purposes
(1) Fluorescence analysis of human/mouse brain slices^[1]:
20-6 μm thick brain slices were incubated with 30 μM C05-05 in 50% (human brain) or 20% (mouse brain) ethanol solution at room temperature for 30 minutes.
Washed with ethanol and ultrapure water in turn, and the slices were sealed.
After sealing, the fluorescence signal of the lesion area was observed by Two-photon laser scanning microscope (excitation wavelength 900 nm, detection wavelength 500-550 nm).

(2) PET imaging of mice^[1]:
Anesthetized mice were intravenously injected with ¹⁸F-C05-05 (24.3±0.7MBq) and immediately scanned for 90-120 min.
Images were reconstructed using Fourier reconstruction and maximum a posteriori probability method, and the distribution of radioactivity in the striatum and midbrain was analyzed.

(3) α-synuclein aggregation inhibition experiment^[1]:
C05-05 (final concentration 20 μM or 200 μM) and protein aggregates were added to the A53T mutant α-synuclein monomer solution (1 mg/mL).
Incubate at 37°C with shaking for 24 h, and then add sarcosine for treatment.
After ultracentrifugation, insoluble aggregates were quantified by electrophoresis and CBB staining.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.