Alpha Synuclein E114C Mutant Pre-formed Fibrils: ATTO 488

Human Recombinant Alpha Synuclein E114C Mutant Pre-formed Fibrils: ATTO 488

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Target: Alpha Synuclein E114C Mutant Pre-formed Fibrils: ATTO 488

Nature: Recombinant

Swiss-Prot: P37840

Expression System: E.coli

Protein Length: 140 aa

Amino Acid Sequence: 10% of mixture (mutant conjugated form): MDVFMKGLSKAKEGVVAAAEKTKQGVAEAAGKTKEGVLYVGSKTKEGVVHGVATVAEKTKEQVTNVGGAVVTGVTAVAQKTVEGAGSIAAATGFVKKDQLGKNEEGAPQEGILCDMPVDPDNEAYEMPSEEGYQDYEPEA90% of mixture (wildtype):MDVFMKGLSKAKEGVVAAAEKTKQGVAEAAGKTKEGVLYVGSKTKEGVVHGVATVAEKTKEQVTNVGGAVVTGVTAVAQKTVEGAGSIAAATGFVKKDQLGKNEEGAPQEGILEDMPVDPDNEAYEMPSEEGYQDYEPEA

Purification: Ion-exchange & SEC purified

Purity: >95%

Storage Buffer: 1X PBS pH 7.4

Protein Size: 14.434 kDa

Conjugate: ATTO 488

Scientific Background: The alpha-synuclein (aSyn) E114C mutation facilitates a single site-specific conjugation with ATTO-488 maleimide that avoids any hindrance on fibrilization or cell entry that may be conferred by non-specific lysine targeting conjugations. This conjugation is ideal due to internal position relative to C-terminal truncation sites, proximity to the NAC, and lack of interference with recruitment in vitro or in primary neurons (1, 2). Pre-formed fibrils (PFFs) generated with 5-25% fluorescently tagged E114C mutants have demonstrated a relative potency >80% compared to wild-type aSyn for inducing misfolding of endogenous aSyn, indicating no significant perturbation of seeding in living cells (1). Atto-488 is a useful tool for identifying cell entry, as the addition of Trypan Blue to cultures prior to imaging will quench fluorescence of extracellular Atto-488 conjugated aSyn (3). Our aSyn E114C-Atto-488 PFFs, which are formed from 10% fluorescently tagged E114C mutants and 90% wild-type monomers, are an excellent tool for studying cell entry and localization, with demonstrated entry into neurons after trypan blue quenching.

References: 1.Haney et al. 2016. Comparison of strategies for non-perturbing labeling of α-synuclein to study amyloidogenesis. Organic & Biomolecular Chemistry. DOI: 10.1039/c5ob02329g2.Karpowicz et al. 2017. Selective imaging of internalized proteopathic a-synuclein seeds in primary neurons reveals mechanistic insight into transmission of synucleinopathies. JBC. DOI: 10.1074/jbc.M117.7802963.Pieri et al. 2016. Structural and functional properties of prefibrillar α-synuclein oligomers. Scientific Reports. DOI: 10.1038/srep24526

Field of Use: Not for use in humans. Not for use in diagnostics or therapeutics. For research use only.

• Artikelnummer: STRSPR-518C-A488
• Hersteller: Stressmarq Biosciences
• Hersteller Artikelnummer: SPR-518C-A488
• Verpackungseinheit: 100 µg x2
• Mengeneinheit: PAK
• Reaktivität: Human
• Methode: Western Blotting, In Vivo Assay