Recombinant mouse Alpha-synuclein protein aggregate (Active) (ab246002)
Key features and details
- Expression system: Escherichia coli
- Purity: > 95% SDS-PAGE
- Active: Yes
- Suitable for: IHC-P, Functional Studies, Electron Microscopy, ICC/IF, SDS-PAGE
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Product name
Recombinant mouse Alpha-synuclein protein aggregate (Active)
See all Alpha-synuclein proteins and peptides -
Biological activity
Endogenous alpha-synuclein phosphorylation. 100 µM alpha synuclein protein monomer seeded with 10 nM alpha synuclein protein PFF (ab246002) in 25 µM Thioflavin T (PBS pH 7.4, 100 µl reaction volume) generated an increased fluorescence intensity after incubation at 37°C with shaking at 600 rpm for 24 hours. Fluorescence was measured by excitation at 450 nm and emission at 485 nm microplate reader.
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Purity
> 95 % SDS-PAGE.
Ion-exchange purified. -
Expression system
Escherichia coli -
Accession
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Protein length
Full length protein -
Animal free
No -
Nature
Recombinant -
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Species
Mouse -
Sequence
MDVFMKGLSKAKEGVVAAAEKTKQGVAEAAGKTKEGVLYVGSKTKEGVVH GVTTVAEKTKEQVTNVGGAVVTGVTAVAQKTVEGAGNIAAATGFVKKDQM GKGEEGYPQEGILEDMPVDPGSEAYEMPSEEGYQDYEPEA -
Predicted molecular weight
15 kDa -
Amino acids
1 to 140 -
Additional sequence information
NP_001035916.1
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Description
Recombinant mouse Alpha-synuclein protein (Active)
Preparation and Storage
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Stability and Storage
Shipped on Dry Ice. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.
Constituent: PBS
This product is an active protein and may elicit a biological response in vivo, handle with caution.
Images
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Immunocytochemistry/ Immunofluorescence - Recombinant mouse Alpha-synuclein protein (Active) (ab246002)
Primary rat hippocampal neurons (DIV16) show lewy body inclusion formation and loss of cells when treated with ab246002 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:proprietary block 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 incluscions. Magnification: 20x.
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Immunohistochemistry (PFA fixed) - Recombinant mouse Alpha-synuclein protein (Active) (ab246002)Immunohistochemistry analysis of rat brain injected with ab246002. Species: Female Sprague-Dawley Rat. Rat was injected with 2µL ab246002 in each of 2 injection sites: AP+1.6, ML+2.4, DV-4.2 from skull; and AP-1.4, ML+0.2, DV-2.8 from skull. 30 days post-injection. Fixation: Saline perfusion followed by 4% PFA fixation for 48 hrs. Secondary Antibody: Biotin-SP Donkey Anti-Rabbit IgG (H+L) at 1:500 for 2 hours in cold room with shaking. ABC signal amplification, DAB staining. Magnification: 20X. Alpha synuclein pathology is seen in the periform/insular cortex and the cingulate cortex on both the same (ipsi) and opposite (contra) sides as the injection sites.
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Electron Microscopy - Recombinant mouse Alpha-synuclein protein (Active) (ab246002)TEM of ab246002. Fibrils were sonicated and image was taken at 100kx magnification.
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Electron Microscopy - Recombinant mouse Alpha-synuclein protein (Active) (ab246002)TEM of ab246002. Image was taken at 100kx magnification.
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Functional Studies - Recombinant mouse Alpha-synuclein protein (Active) (ab246002)ab246002 seeds the formation of new Alpha synuclein fibrils from the pool of active alpha synuclein monomers. 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 nM of ab246002 is combined with 100 µM of active Alpha synuclein monomer, as compared to ab246002 and active alpha Synuclein monomer alone. Thioflavin T ex = 450 nm, em = 485 nm.
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SDS-PAGE - Recombinant mouse Alpha-synuclein protein (Active) (ab246002)SDS-PAGE analysis of ab246002 (2 μg).
