Alpha Synuclein Protein

Human Recombinant Alpha Synuclein Oligomers (Kinetically Stable)

Catalog No. SPR-484

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Expression System E. coli
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SPR-484_Alpha-Synuclein-Oligomers-Kinetcially-Stable-Protein-TEM-1.png
Native PAGE of Human Recombinant Alpha Synuclein Oligomers (Kinetically Stable) (SPR-484).Αlpha Synuclein Oligomers Have Distinct Secondary Structure Differences Compared to Fibrils.Toxicity of Human Recombinant Alpha Synuclein Oligomers (Kinetically Stable) (SPR-484).IHC of Human Recombinant Alpha Synuclein Oligomers (Kinetically Stable) (SPR-484).
Product Name Alpha Synuclein Protein
Description

Human Recombinant Alpha Synuclein Oligomers (Kinetically Stable)
Applications WB, Native PAGE, In vivo assay, In vitro assay
Concentration 2 mg/mL
Conjugates No tag
Dylight 488
Overview:

  • High fluorescence yield
  • High photostability
  • Less pH-sensitive
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested
  • Molecular weight: 1011 g/mol

Dylight 488 Datasheet

 Dylight 488 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 493 nm

λem = 518 nm

εmax = 7.0×104

Laser = 488 nm

 

APC/Cy7
Overview:

  • High quantum yield
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested

APC-Cy7 Datasheet

 

 ACP-Cy7 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 652 nm

λem = 790 nm

Laser = 594 or 633 nm

 

 

  Dylight 350
Overview:

  • High fluorescence intensity
  • High photostability
  • Less pH-sensitive
  • Excellent solubility in water
  • Stringently QC tested
  • Excellent batch-to-batch reproducibility
  • Molecular weight: 874 g/mol

Dylight 350 Datasheet

 Dylight 350 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 353 nm

λem = 432 nm

εmax = 1.5×104

 

 

  Dylight 405
Overview:

  • High fluorescence intensity
  • High photostability
  • Less pH-sensitive
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested
  • Molecular weight: 793 g/mol

Dylight 405 Datasheet

 Dylight 405 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 400 nm

λem = 420 nm

εmax = 3.0×104

Laser = 405 nm

 

Dylight 594
Overview:

  • High fluorescence yield
  • High photostability
  • Less pH-sensitive
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested
  • Molecular weight: 1078 g/mol

Dylight 594 Datasheet

 Dylight 594 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 593 nm

λem = 618 nm

εmax = 8.0×104

Laser = 526 nm

 

 Dylight 633
Overview:

  • High fluorescence yield
  • High photostability
  • Less pH-sensitive
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested
  • Molecular weight: 1066 g/mol

Dylight 633 Datasheet

 Dylight 633 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 638 nm

λem = 658 nm

εmax = 1.7×105

Laser = 633 nm

 

 PerCP 
Overview:

  • Peridinin-Chlorophyll-Protein Complex
  • Small phycobiliprotein
  • Isolated from red algae
  • Large stokes shift (195 nm)
  • Molecular Weight: 35 kDa

PerCP Datasheet

  PerCP Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 482 nm

λem = 677 nm

εmax = 1.96 x 106

Laser = 488 nm

 

 PE/ATTO 594
PE/ATTO 594 is a tandem conjugate, where PE is excited at 535 nm and transfers energy to ATTO 594 via FRET (fluorescence resonance energy transfer), which emits at 627 nm.
Overview:

  • High fluorescence yield
  • High photostability
  • Very hydrophilic
  • Excellent solubility in water
  • Very little aggregation

PE/ATTO 594 Datasheet

 PE-ATTO 594 Fluorophore Conjugate Excitation and Emission Spectra Optical Properties:

λex = 535 nm

λem = 627 nm

Laser = 488 to 561 nm

 

  FITC (Fluorescein)
Overview:

  • Excellent fluorescence quantum yield
  • High rate of photobleaching
  • Good solubility in water
  • Broad emission spectrum
  • pH dependent spectra
  • Molecular formula: C20H12O5
  • Molar mass: 332.3 g/mol

FITC-Fluorescent-conjugate

 FITC Fluorescein Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 494 nm

λem = 520 nm

εmax = 7.3×104

Φf = 0.92

τfl = 5.0 ns

Brightness = 67.2

Laser = 488 nm

Filter set = FITC

 

 ATTO 700
Overview:

  • High fluorescence yield
  • Excellent thermal and photostability
  • Quenched by electron donors
  • Very hydrophilic
  • Good solubility in polar solvents
  • Zwitterionic dye
  • Molar Mass: 575 g/mol

ATTO 700 Datasheet

  ATTO 700 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 700 nm

λem = 719 nm

εmax = 1.25×105

Φf = 0.25

τfl = 1.6 ns

Brightness = 31.3

Laser = 676 nm

Filter set = Cy®5.5

 

 ATTO 680
Overview:

  • High fluorescence yield
  • Excellent thermal and photostability
  • Quenched by electron donors
  • Very hydrophilic
  • Good solubility in polar solvents
  • Zwitterionic dye
  • Molar Mass: 631 g/mol

ATTO 680 Datasheet

  ATTO 680 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 680 nm

λem = 700 nm

εmax = 1.25×105

Φf = 0.30

τfl = 1.7 ns

Brightness = 37.5

Laser = 633 – 676 nm

Filter set = Cy®5.5

 

 ATTO 655
Overview:

  • High fluorescence yield
  • High thermal and photostability
  • Excellent ozone resistance
  • Quenched by electron donors
  • Very hydrophilic
  • Good solubility in polar solvents
  • Zwitterionic dye
  • Molar Mass: 634 g/mol

ATTO 655 Datasheet

 ATTO 655 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 663 nm

λem = 684 nm

εmax = 1.25×105

Φf = 0.30

τfl = 1.8 ns

Brightness = 37.5

Laser = 633 – 647 nm

Filter set = Cy®5

 

 ATTO 633
Overview:

  • High fluorescence yield
  • High thermal and photostability
  • Moderately hydrophilic
  • Good solubility in polar solvents
  • Stable at pH 4 – 11
  • Cationic dye, perchlorate salt
  • Molar Mass: 652.2 g/mol

ATTO 633 Datasheet

 ATTO 633 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 629 nm

λem = 657 nm

εmax = 1.3×105

Φf = 0.64

τfl = 3.2 ns

Brightness = 83.2

Laser = 633 nm

Filter set = Cy®5

 

 ATTO 594
Overview:

  • High fluorescence yield
  • High photostability
  • Very hydrophilic
  • Excellent solubility in water
  • Very little aggregation
  • New dye with net charge of -1
  • Molar Mass: 1137 g/mol

ATTO 594 Datasheet

  ATTO 594 Fluorophore Excitation and Emission Spectrum Optical Properties:

λex = 601 nm

λem = 627 nm

εmax = 1.2×105

Φf = 0.85

τfl = 3.5 ns

Brightness = 102

Laser = 594 nm

Filter set = Texas Red®

 

 ATTO 565
Overview:

  • High fluorescence yield
  • High thermal and photostability
  • Good solubility in polar solvents
  • Excellent solubility in water
  • Very little aggregation
  • Rhodamine dye derivative
  • Molar Mass: 611 g/mol

ATTO 565 Datasheet

  ATTO 565 Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 563 nm

λem = 592 nm

εmax = 1.2×105

Φf = 0.9

τfl = 3.4 n

Brightness = 10

Laser = 532 nm

Filter set = TRITC

 

  ATTO 488
Overview:

  • High fluorescence yield
  • High photostability
  • Very hydrophilic
  • Excellent solubility in water
  • Very little aggregation
  • New dye with net charge of -1
  • Molar Mass: 804 g/mol 

ATTO 488 Datasheet

   ATTO 488 Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 501 nm

λem = 523 nm

εmax = 9.0×104

Φf = 0.80

τfl = 4.1 ns

Brightness = 72

Laser = 488 nm

Filter set = FITC

 

  ATTO 390
Overview:

  • High fluorescence yield
  • Large Stokes-shift (89 nm)
  • Good photostability
  • Moderately hydrophilic
  • Good solubility in polar solvents
  • Coumarin derivate, uncharged
  • Low molar mass: 343.42 g/mol 

ATTO 390 Datasheet

 ATTO 390 Fluorescent Dye Excitation and Emission Spectra Optical Properties:

λex = 390 nm

λem = 479 nm

εmax = 2.4×104

Φf = 0.90

τfl = 5.0 ns

Brightness = 21.6

Laser = 365 or 405 nm

 

APC (Allophycocyanin)
Overview:

  • High quantum yield
  • Large phycobiliprotein
  • 6 chromophores per molecule
  • Isolated from red algae
  • Molecular Weight: 105 kDa

APC Datasheet

  APC Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 650 nm

λem = 660 nm

εmax = 7.0×105

Φf = 0.68

Brightness = 476

Laser = 594 or 633 nm

Filter set = Cy®5

 

Streptavidin

Properties:

  • Homo-tetrameric protein purified from Streptomyces avidinii which binds four biotin molecules with extremely high affinity
  • Molecular weight: 53 kDa
  • Formula: C10H16N2O3S
  • Applications: Western blot, immunohistochemistry, and ELISA

Streptavidin Datasheet

BiotinBiotin Conjugate Structure

Properties:

  • Binds tetrameric avidin proteins including Streptavidin and neuravidin with very high affinity
  • Molar mass: 244.31 g/mol
  • Formula: C10H16N2O3S
  • Applications: Western blot, immunohistochemistry, and ELISA

Biotin Datasheet

HRP (Horseradish peroxidase)

Properties:

  • Enzymatic activity is used to amplify weak signals and increase visibility of a target
  • Readily combines with hydrogen peroxide (H2O2) to form HRP-H2O2 complex which can oxidize various hydrogen donors
  • Catalyzes the conversion of:
    • Chromogenic substrates (e.g. TMB, DAB, ABTS) into coloured products
    • Chemiluminescent substrates (e.g. luminol and isoluminol) into light emitting products via enhanced chemiluminescence (ECL)
    • Fluorogenic substrates (e.g. tyramine, homovanillic acid, and 4-hydroxyphenyl acetic acid) into fluorescent products
  • High turnover rate enables rapid generation of a strong signal
  • 44 kDa glycoprotein
  • Extinction coefficient: 100 (403 nm)
  • Applications: Western blot, immunohistochemistry, and ELISA

HRP Datasheet

AP (Alkaline Phosphatase)

Properties:

  • Broad enzymatic activity for phosphate esters of alcohols, amines, pyrophosphate, and phenols
  • Commonly used to dephosphorylate the 5’-termini of DNA and RNA to prevent self-ligation
  • Catalyzes the conversion of:
    • Chromogenic substrates (e.g. pNPP, naphthol AS-TR phosphate, BCIP) into coloured products
    • Fluorogenic substrates (e.g. 4-methylumbelliferyl phosphate) into fluorescent products
  • Molecular weight: 140 kDa
  • Applications: Western blot, immunohistochemistry, and ELISA

AP Datasheet

  R-PE (R-Phycoerythrin)
Overview:

  • Broad excitation spectrum
  • High quantum yield
  • Photostable
  • Member of the phycobiliprotein family
  • Isolated from red algae
  • Excellent solubility in water
  • Molecular Weight: 250 kDa

R-PE Datasheet

  R-PE Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 565 nm

λem = 575 nm

εmax = 2.0×106

Φf = 0.84

Brightness = 1.68 x 103

Laser = 488 to 561 nm

Filter set = TRITC

 
Nature Recombinant
Species Human
Expression System E. coli
Amino Acid Sequence MDVFMKGLSK AKEGVVAAAE KTKQGVAEAA GKTKEGVLYV GSKTKEGVVH GVATVAEKTK EQVTNVGGAV VTGVTAVAQK TVEGAGSIAA ATGFVKKDQL GKNEEGAPQE GILEDMPVDP DNEAYEMPSE EGYQDYEPEA
Purity >95%
Other Resources
Protein Length Full Length
Protein Size 650-1200 kDa
Field of Use Not for use in humans. Not for use in diagnostics or therapeutics. For in vitro research use only.

Properties

Storage Buffer PB pH 7.4 (10 mM KH2PO4, 7.5 mM NaOH, pH 7.4)
Storage Temperature -80ºC
Shipping Temperature Dry Ice. Shipping note: Product will be shipped separately from other products purchased in the same order.
Purification Ion-exchange Purified, monomer removed with 100K MWCO filter
Cite This Product Human Recombinant Alpha Synuclein Oligomers (StressMarq Biosciences Inc., Victoria BC CANADA, Catalog # SPR-484)
Certificate of Analysis Certified >95% pure using SDS-PAGE analysis. Low endotoxin <5 EU/mL @ 2mg/mL.
Other Relevant Information Kinetically stable alpha synuclein oligomers are generated from monomer without an inducer/inhibitor, remain soluble oligomers for at least two weeks at 37 degrees, and are toxic to dopaminergic neurons.

Biological Description

Alternative Names Alpha synuclein protein, Alpha-synuclein oligomer, Alpha synuclein protein oligomer, Alpha-synuclein protein, Non-A beta component of AD amyloid protein, Non-A4 component of amyloid precursor protein, NACP protein, SNCA protein, NACP protein, PARK1 protein, Alpha synuclein oligomers, Alpha Synuclein Protein Oligomers, SYN protein, Parkinson's disease familial 1 Protein
Research Areas Alzheimer's Disease, Neurodegeneration, Neuroscience, Parkinson's Disease, Synuclein, Tangles & Tau, Multiple System Atrophy
Cellular Localization Cell membrane, Cytoplasm, Nucleus, Presynaptic Termini
Accession Number NP_000336.1
Gene ID 6622
Swiss Prot P37840
Scientific Background Our kinetically stable oligomers of alpha-synuclein are generated without an inducer or inhibitor and remain stable for at least 2 weeks at 37oC. They present as globular structures under TEM, demonstrate toxicity in rat primary dopaminergic neurons and induce Parkinson’s-associated alpha synuclein phosphoserine 129 pathology. These oligomers have been previously characterized as globular, cylindrical structures with a beta-sheet structure intermediate between monomers and fibrils, and were demonstrated to have a higher toxicity to neurons than alpha-synuclein fibrils (1,2). Alpha-Synuclein (SNCA) is expressed predominantly in the brain, where it is concentrated in presynaptic nerve terminals (3). Alpha-synuclein is highly expressed in the mitochondria of the olfactory bulb, hippocampus, striatum and thalamus (4). Functionally, it has been shown to significantly interact with tubulin (5), and may serve as a potential microtubule-associated protein. It has also been found to be essential for normal development of the cognitive functions; inactivation may lead to impaired spatial learning and working memory (6). SNCA fibrillar aggregates represent the major non A-beta component of Alzheimer’s disease amyloid plaque, and a major component of Lewy body inclusions, and Parkinson's disease. Parkinson's disease (PD) is a common neurodegenerative disorder characterized by the progressive accumulation in selected neurons of protein inclusions containing alpha-synuclein and ubiquitin (7, 8).
References 1. Chen, S.W., et. al. (2015). PNAS. E1994-E2003.
2. Lorenzen, N., et. al. (2014). JACS. 136: 3859-3868.
3. “Genetics Home Reference: SNCA”. US National Library of Medicine. (2013).
4. Zhang L., et al. (2008) Brain Res. 1244: 40-52.
5. Alim M.A., et al. (2002) J Biol Chem. 277(3): 2112-2117.
6. Kokhan V.S., Afanasyeva M.A., Van'kin G. (2012) Behav. Brain. Res. 231(1): 226-230.
7. Spillantini M.G., et al. (1997) Nature. 388(6645): 839-840.
8. Mezey E., et al. (1998) Nat Med. 4(7): 755-757.

Product Images

<p>TEM of kinetically stable alpha-synuclein oligomers (SPR-484). Negative stain transmission electron microscopy images of SPR-484 acquired at 80 Kv on carbon coated 400 mesh copper grids using phosphotungstic acid and uranyl acetate stain.</p>

TEM of kinetically stable alpha-synuclein oligomers (SPR-484). Negative stain transmission electron microscopy images of SPR-484 acquired at 80 Kv on carbon coated 400 mesh copper grids using phosphotungstic acid and uranyl acetate stain.

<p>Kinetically stable alpha-synuclein oligomers (SPR-484) are toxic to dopaminergic neurons and induce phosphorylation of alpha-synuclein Ser129, a pathology associated with Parkinson’s disease. Survival of rat primary dopaminergic neurons 11 days after treatment quantified by anti-MAP2 antibody and expressed as a percentage of control (A). Levels of alpha-synuclein pSer129 present in rat primary dopaminergic neurons 11 days after treatment quantified by ratio of anti-alpha-synuclein pSer129 antibody to anti-MAP2 antibody expressed as a percentage of control (B). Mean +/- s.e.m; ** p<0.01 stats vs control, one-way Anova followed by Dunnett’s test; § p<0.05, stats vs control, Student’s t-test. Data is representative of n=6 experimental repeats for each condition; # represents control, n.s. indicates not significant p>0.05.</p>

Kinetically stable alpha-synuclein oligomers (SPR-484) are toxic to dopaminergic neurons and induce phosphorylation of alpha-synuclein Ser129, a pathology associated with Parkinson’s disease. Survival of rat primary dopaminergic neurons 11 days after treatment quantified by anti-MAP2 antibody and expressed as a percentage of control (A). Levels of alpha-synuclein pSer129 present in rat primary dopaminergic neurons 11 days after treatment quantified by ratio of anti-alpha-synuclein pSer129 antibody to anti-MAP2 antibody expressed as a percentage of control (B). Mean +/- s.e.m; ** p<0.01 stats vs control, one-way Anova followed by Dunnett’s test; § p<0.05, stats vs control, Student’s t-test. Data is representative of n=6 experimental repeats for each condition; # represents control, n.s. indicates not significant p>0.05.

<p>Representative immunohistochemistry images of Parkinson’s-associated pSer129 pathology induced in rat primary dopaminergic cells by kinetically stable alpha-synuclein oligomers (SPR-484). Primary rat dopaminergic neurons 11 days after treatment with control PBS buffer (A). Primary rat dopaminergic neurons 11 days after treatment with 10 µg/mL SPR-484 (B). Nuclei appear blue (Hoechst), dopaminergic neurons appear red (MAP2) and pathology appears green (α-syn pSer129). Both cultures treated with chicken polyclonal anti-MAP-2 antibody, mouse monoclonal anti-αsyn pSer129-specific antibody, Alexa Fluor 488 goat anti-mouse IgG, Alexa Fluor 647 goat anti-chicken IgG, and fluorescent marker Hoechst in the same solution. White arrows emphasize several regions of strong presence of pSer129 pathology in MAP2 positive neurons. Scale bar represents 100 μm. Note: SPR-484 is generated recombinantly in E.coli and is non-phosphorylated prior to addition on neurons.</p>

Representative immunohistochemistry images of Parkinson’s-associated pSer129 pathology induced in rat primary dopaminergic cells by kinetically stable alpha-synuclein oligomers (SPR-484). Primary rat dopaminergic neurons 11 days after treatment with control PBS buffer (A). Primary rat dopaminergic neurons 11 days after treatment with 10 µg/mL SPR-484 (B). Nuclei appear blue (Hoechst), dopaminergic neurons appear red (MAP2) and pathology appears green (α-syn pSer129). Both cultures treated with chicken polyclonal anti-MAP-2 antibody, mouse monoclonal anti-αsyn pSer129-specific antibody, Alexa Fluor 488 goat anti-mouse IgG, Alexa Fluor 647 goat anti-chicken IgG, and fluorescent marker Hoechst in the same solution. White arrows emphasize several regions of strong presence of pSer129 pathology in MAP2 positive neurons. Scale bar represents 100 μm. Note: SPR-484 is generated recombinantly in E.coli and is non-phosphorylated prior to addition on neurons.

<p>Αlpha Synuclein Oligomers Have Distinct Secondary Structure Differences Compared to Fibrils. UV-CD data suggests that StressMarq’s Alpha Synuclein Oligomers have distinct secondary structure differences compared to our monomers and fibrils. More specifically, StressMarq’s Kinetically Stable Alpha Synuclein Oligomers (SPR-484) show a significantly higher alpha helix content and lower beta sheet/turn content than our Alpha Synuclein Pre-formed Fibrils (Type 1) (SPR-322). StressMarq’s Alpha Synuclein Monomers (SPR-316) show a strong negative signal at 200 nm indicative of a disordered protein state (low secondary structure content).</p>

Αlpha Synuclein Oligomers Have Distinct Secondary Structure Differences Compared to Fibrils. UV-CD data suggests that StressMarq’s Alpha Synuclein Oligomers have distinct secondary structure differences compared to our monomers and fibrils. More specifically, StressMarq’s Kinetically Stable Alpha Synuclein Oligomers (SPR-484) show a significantly higher alpha helix content and lower beta sheet/turn content than our Alpha Synuclein Pre-formed Fibrils (Type 1) (SPR-322). StressMarq’s Alpha Synuclein Monomers (SPR-316) show a strong negative signal at 200 nm indicative of a disordered protein state (low secondary structure content).

<p>Kinetically stable alpha-synuclein oligomers (SPR-484) are stable after a freeze-thaw cycle and when incubated at 37 oC for 2 weeks. Tris-Gylcine Native PAGE migration of kinetically stable alpha-synuclein oligomers after a freeze-thaw cycle (A). Size-exclusion chromatography of SPR-484 after a freeze-thaw cycle (B) and 37oC incubation for 14 days (C). By peak area, approximately 90% of SPR-484 remains oligomeric after freeze-thaw and 37oC treatments. SEC was performed on Superdex 200 10/300 GL Increase column in phosphate buffer pH 7.4. Note: Monomeric alpha-synuclein is an intrinsically disordered 14 kDa protein. Due to its extended conformation in solution, migration of free monomeric alpha-synuclein is similar to that of a globular 60 kDa protein on Native PAGE and SEC.</p>

Kinetically stable alpha-synuclein oligomers (SPR-484) are stable after a freeze-thaw cycle and when incubated at 37 oC for 2 weeks. Tris-Gylcine Native PAGE migration of kinetically stable alpha-synuclein oligomers after a freeze-thaw cycle (A). Size-exclusion chromatography of SPR-484 after a freeze-thaw cycle (B) and 37oC incubation for 14 days (C). By peak area, approximately 90% of SPR-484 remains oligomeric after freeze-thaw and 37oC treatments. SEC was performed on Superdex 200 10/300 GL Increase column in phosphate buffer pH 7.4. Note: Monomeric alpha-synuclein is an intrinsically disordered 14 kDa protein. Due to its extended conformation in solution, migration of free monomeric alpha-synuclein is similar to that of a globular 60 kDa protein on Native PAGE and SEC.

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