Alpha Synuclein A90C Mutant Monomers

Human Recombinant Alpha Synuclein A90C Mutant Monomers

Product Specifications

Background

Thioflavin-T (ThT) fluorescence remains a common measurement of alpha-synuclein fibril formation, yet ThT exhibits poor affinity for oligomers and early aggregates. The alpha-synuclein A90C mutant monomers can be specifically labelled with alternative fluorophores (such as Alexa 488/Alexa 647) via maleimide chemistry to enable more sensitive FRET analysis of aggregation. The A90C mutant showed no perturbation of monomer structure and Alexa Fluor dye attachment to cysteine 90 was demonstrated to have no effect on the kinetics of fibril formation (1-3) . Residue 90 is at the periphery of the NAC region, a key constituent of the alpha-synuclein β-sheet fibril core, which results in fluorophores on different monomers coming into close proximity upon formation of β-sheet structure during aggregation (4) . Note - to prevent the potential mis-translation of alpha-synuclein Y136 as C136 during E.coli expression, the Y136-TAT construct was used (5) .

Product Name Alternative

SNCA, alpha-synuclein, synuclein, Alpha synuclein monomer, Alpha-synuclein monomer, Alpha synuclein protein monomer, Alpha synuclein monomer, Alpha-synuclein protein, SNCA protein

UNSPSC

12352202

UN Code

Non-hazardous

Hazard Statement

Non-hazardous

Swiss Prot

P37840-1 (wildtype)

Expression System

E. coli

Host

E. coli

Origin Species

Human

Target

Alpha Synuclein A90C

Conjugation

No Tag

Nature

Recombinant

Sequence

MDVFMKGLSKAKEGVVAAAEKTKQGVAEAAGKTKEGVLYVGSKTKEGVVHGVATVAEKTKEQVTNVGGAVVTGVTAVAQKTVEGAGSIACATGFVKKDQLGKNEEGAPQEGILEDMPVDPDNEAYEMPSEEGYQDYEPEA

Applications

WB | SDS PAGE | In vitro Assay | Conjugation | FRET

Field of Research

Purification Method

Ion-exchange Purified

Purification

Ion-exchange Purified

Limit Of Detection

Protein certified >95% pure on SDS-PAGE & Nanodrop analysis

Concentration

2 mg/ml

Purity

>95%

Weight

0.02

Length

Full length (1 - 140 aa)

Buffer

20mM Hepes pH 7.4, 150mM NaCl, 1mM TCEP pH 7.0

Molecular Weight

14.49 kDa

Precautions

Not for use in humans. Not for use in diagnostics or therapeutics. For research use only.

Additionnal Information

TCEP present to prevent disulfide bonds. Maleimide coupling reactions can be performed efficiently in the presence of TCEP.

References & Citations

1.Thirunavukkuarasu, et al. 2008. Multiparametric Flurorescence Detection of Early Stages in the Amylord Protein Aggregation of Pyrene-labeled α-Synuclein. J. Mol. Biol. 378(5): 1064-73. https://doi.org/10.1016/j.jmb.2008.03.034 2.Cremades, et al. 2012. Direct Observation of the Interconversion of Normal and Toxic Forms of α-Synuclein. Cell. 149(5): 1048-59. doi: 10.1016/j.cell.2012.03.037 3.Horrocks et al. 2015. Fast Flow Microfluidics and Single-Molecule Fluorescence for the Rapid Characterization of α-Synuclein Oligomers. Anal. Chem. 87(17): 8818-26. https://doi.org/10.1021/acs.analchem.5b01811 4.Iljina et al. 2016. Kinetic model of the aggregation of alpha-synuclein provides insights into prion-like spreading. PNAS. 113(19): E1206-15. doi: 10.1073/pnas.1524128113 5.Masuda, et al. 2006. Cysteine misincorporation in bacterially expressed human alpha-synuclein. FEBS Lett. 580(7): 1775-9. doi: 10.1016/j.febslet.2006.02.032

Shipping Conditions

Dry Ice. Shipping note: Product will be shipped separately from other products purchased in the same order.

Storage Conditions

-80ºC

Notes

TCEP present to prevent disulfide bonds. Maleimide coupling reactions can be performed efficiently in the presence of TCEP.

Protein Length

Full length (1 - 140 aa)

Background Reference 01

1.Thirunavukkuarasu, et al. 2008. Multiparametric Flurorescence Detection of Early Stages in the Amylord Protein Aggregation of Pyrene-labeled α-Synuclein. J. Mol. Biol. 378 (5) : 1064-73. https://doi.org/10.1016/j.jmb.2008.03.034 2.Cremades, et al. 2012. Direct Observation of the Interconversion of Normal and Toxic Forms of α-Synuclein. Cell. 149 (5) : 1048-59. doi: 10.1016/j.cell.2012.03.037 3.Horrocks et al. 2015. Fast Flow Microfluidics and Single-Molecule Fluorescence for the Rapid Characterization of α-Synuclein Oligomers. Anal. Chem. 87 (17) : 8818-26. https://doi.org/10.1021/acs.analchem.5b01811 4.Iljina et al. 2016. Kinetic model of the aggregation of alpha-synuclein provides insights into prion-like spreading. PNAS. 113 (19) : E1206-15. doi: 10.1073/pnas.1524128113 5.Masuda, et al. 2006. Cysteine misincorporation in bacterially expressed human alpha-synuclein. FEBS Lett. 580 (7) : 1775-9. doi: 10.1016/j.febslet.2006.02.032