C57/BL6 mice were injected with sonicated recombinant mouse alpha synuclein monomers or fibrils at 8 weeks of age. Mice were unilaterally injected in the dorsal striatum (bregma AP + 0.2 mm, L +/1 2.0 mm, V - 3.0 mm) and sacrificed 30 days post-injection. (A) 1.25 uL mouse alpha synuclein monomers (SPR-323). (B) 2.5 uL mouse alpha synuclein monomers (SPR-323). (C) 2.5 ug alpha synuclein PFFs (SPR-324). (C) 5 ug alpha synuclein PFFs (SPR-324) Inset: PBS (negative control). Primary antibody: Anti-Alpha Synuclein pSer129 (SMC-600) at 1:10 000. Secondary antibody: anti-rabbit HRP. Mice injected with PFF displayed alpha synuclein staining in the striatum and cortex and contralateral to the injection site. Courtesy of: Porsolt.
Type 1 alpha synuclein pre-formed fibrils (SPR-324) seed the formation of new alpha synuclein fibrils from the pool of alpha synuclein monomers (SPR-323). Thioflavin T is a fluorescent dye that binds to beta sheet-rich structures, such as those in alpha synuclein fibrils. Upon binding, the emission spectrum of the dye experiences a red-shift, and increased fluorescence intensity. Thioflavin T emission curves show increased fluorescence (correlated to alpha synuclein protein aggregation) over time when 10 µM of Type 1 alpha synuclein pre-formed fibrils (SPR-324) is combined with 100 µM of alpha synuclein monomer (SPR-323), as compared to Type 1 alpha synuclein pre-formed fibrils (SPR-324) or alpha synuclein monomer (SPR-323) alone. Thioflavin T ex = 450 nm, em = 485 nm.
SDS-PAGE of ~14 kDa Mouse Recombinant Alpha Synuclein Protein Monomer (SPR-323). Lane 1: Molecular Weight Ladder (MW). Lane 2: Alpha Synuclein Protein Monomer (2 µg) (SPR-323).
Supplier Page from StressMarq Biosciences Inc for Alpha Synuclein Monomers