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Proteins and Peptides

Tau-441 (2N4R) and Alpha Synuclein Co-Polymer Fibrils

Product Sizes
100 ug
SPR-495-100UG
2 x 100 ug
SPR-495-2X100UG
5 x 100 ug
SPR-495-5X100UG
About this Product
SKU:
SPR-495
Additional Names:
MAPT, 2N4R, Tau40 neurofibrillary tangle protein, paired-helical filament, PHFs, SNCA, NACP, PARK1, asyn, alpha-synuclein, pre-formed fibril, PFFs, mixed fibrils
Application:
Cell-based/Functional Assay, SDS-PAGE, Western Blot
CE/IVD:
RUO
Concentration:
2 mg/ml
Extra Details:
Brain-specific tau isoforms vary in the number of N-terminal inserts and C- terminal repeat domains due to alternative splicing of exons; the 2N4R isoform of tau is expressed in adult brain yet is absent from the fetal brain (1). Tau and alpha-synuclein polymerize into amyloid fibrils to form filamentous inclusions in neurodegenerative diseases such as Alzheimer's and Parkinson's disease. Tau has been shown to interact with alpha-synuclein in vitro (2), with synergistic cross-seeding between tau and alpha-synuclein resulting in polymerization of each other into fibrillary amyloid lesions in neuronal cultures and in vivo (3,4). Recombinant tau and alpha-synuclein co-polymer fibrils have demonstrated a more widespread transmission of induced pathology in a rodent model of tauopathies compared to pure Tau or alpha-synuclein fibrils alone (5). These co-polymer fibrils have also shown enhanced alpha-synuclein aggregation in vitro, and more severe alpha-synuclein pathology and Parkinson's disease-like symptoms in mice (6).
Immunogen:
Tau and Alpha Synuclein Co-Polymer
Molecular Weight:
2N4R: 45.84 kDa, ASYN: 14.46 kDa
Purity:
>95%
Purification:
Ion Exchange
Sequence:
Tau: MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKESPLQTPTEDGSEEPGSETSDAKSTPTAEDVTAPLVDEGAPGKQAAAQPHTEIPEGTTAEEAGIGDTPSLEDEAAGHVTQARMVSKSKDGTGSDDKKAKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPTREPKKVAVVRTPPKSPSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIINKKLDLSNVQSKCGSKDNIKHVPGGGSVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL Asyn: MDVFMKGLSKAKEGVVAAAEKTKQGVAEAAGKTKEGVLYVGSKTKEGVVHGVATVAEKTKEQVTNVGGAVVTGVTAVAQKTVEGAGSIAAATGFVKKDQLGKNEEGAPQEGILEDMPVDPDNEAYEMPSEEGYQDYEPEA
Shipping Conditions:
Dry Ice
Storage Conditions:
-70[o]C
Supplier:
StressMarq Biosciences
Type:
Proteins, Peptides, Small Molecules & Other Biomolecules: Recombinant Proteins
TEM of Tau 2N4R & Alpha Synuclein Co-Polymer Fibrils (SPR-495). Negative stain transmission electron microscopy images of SPR-495 acquired at 80 Kv on carbon coated 400 mesh copper grids using phosphotungstic acid and uranyl acetate stain.
Immuno-TEM of Tau & Alpha Synuclein Co-Polymer Fibrils. Fibrils absorbed onto carbon- coated copper grids. Grids were blocked with 1% BSA and 0.1% Tween-20 in PBS, incubated sequentially with primary antibodies at 20 ug/mL in blocking buffer, washed, then incubated with secondary antibodies at 20 ug/mL in blocking buffer, washed and stained with 2% uranyl acetate as a negative stain. 6nm (asyn) and 12nm (tau) signals only appear together in the same fibril strand for the co-polymer fibril samples, and no antibody cross reactivity is observed in control alpha-synuclein or tau fibrils. Primary antibodies: Mouse anti-human asyn monoclonal (StressMarq SMC-532) and rabbit anti-human tau polyclonal (StressMarq SPC-802); Secondary antibodies: 6nm Colloidal Gold-conjugated Goat Anti-Mouse (Jackson ImmunoResearch 115-195- 146) and 12nm Colloidal Gold-conjugated Goat Anti-Rabbit (Jackson ImmunoResearch 111-205-144).
Bis-Tris SDS-PAGE (12%) of Tau 2N4R & Alpha Synuclein Co-Polymer Fibrils. MW ladder = Precision Plus Protein All Blue pre-stained standards.
In vitro Seeding activity of Tau 2N4R & Alpha Synuclein Co-Polymer Fibrils in ThT Assay. Tau 2N4R and Alpha Synuclein co-polymer fibrils seed fibril formation of both Alpha Synuclein monomers and of a mixture of Alpha Synuclein and Tau 2N4R monomers over 72 hours. Reactions (100uL) shaken at 600 rpm in Greiner-Bio 96 Well Non-Binding Cell Culture Microplates, Black (Greiner-Bio Catalog #655900) at 37°C and read with an XPS Microplate Reader set at 450nmex/485nmem.