Target: Alpha Synuclein E114C Mutant Monomers: ATTO 488
Nature: Recombinant
Swiss-Prot: P37840
Expression System: E.coli
Protein Length: 140 aa
Amino Acid Sequence: MDVFMKGLSKAKEGVVAAAEKTKQGVAEAAGKTKEGVLYVGSKTKEGVVHGVATVAEKTKEQVTNVGGAVVTGVTAVAQKTVEGAGSIAAATGFVKKDQLGKNEEGAPQEGILCDMPVDPDNEAYEMPSEEGYQDYEPEA
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 contain 10% fluorescently tagged E114C mutants, 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 in vitro research use only.
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