Primary rat hippocampal neurons show lewy body inclusion formation and loss of cells when treated with Type 1 mouse Alpha Synuclein Protein Pre-formed Fibrils (SPR-324) at 4 µg/ml (D-F) on DVI2, but not when treated with a control (A-C). Tissue: Primary hippocampal neurons. Species: Sprague-Dawley rat. Fixation: 3% formaldehyde from PFA for 20 min. Blocker: 1:1 PBS:LiCOR Odyssey Block (LiCOR, 927-40010) and 30 mL/mL of 0.1% triton-X 100 for 30 min. Primary Antibody: Mouse anti-pSer129 Antibody (1:1000) and Rabbit anti-pSer129 (1:800) for 24 hours at 4°C. Secondary Antibody: ATTO 546 Donkey Anti-Mouse (1:700) and ATTO 488 Donkey Anti-Rabbit (1:700) for 1 hour at RT (composite green). Counterstain: Hoechst (blue) nuclear stain at 1:3000 for 1 hour at RT. Localization: Lewy body inclusions. Magnification: 20x.
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). (D) 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.
C57/BL6 mice were injected with 5 ug sonicated mouse recombinant alpha synuclein PFFs (SPR-324) 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) contralateral cortex. (B) ipsilateral cortex. (C) contralateral striatum. (D) ipsilateral striatum. 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.
Primary mouse hippocampal neurons treated with 100 nM sonicated mouse alpha synuclein PFFs (SPR-324) (A). Phosphorylated alpha synuclein (detected with pSer129 antibody SPC-742) was visible in perinucleus and neurites compared to untreated control (B). Read the protocol at pabmabs.com/?p=7901. Image courtesy of Trine Rasmussen, Simon Molgaard Jensen at Aarhus University.
TEM of Type 1 mouse alpha synuclein Pre-formed Fibrils (SPR-324). Image was taken at 100kx magnification.
Supplier Page from StressMarq Biosciences Inc for Alpha Synuclein Pre-formed Fibrils