Alpha Synuclein Pre-formed Fibrils: Biotinylated

Cynomolgus Recombinant Alpha Synuclein PFFs: Biotinylated (C-terminus)

Catalog No. SPR-528

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Expression System E. coli
Tags C-terminal 15 amino acid tag: biotinylated
SKU: SPR-528 Category:

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SPR-528_Alpha-Synuclein-Pre-formed-Fibrils-Biotinylated-Protein-SDS-Page-1.png
TEM of Cynomolgus Alpha Synuclein Pre-formed Fibrils: Biotinylated Protein (SPR-528)Sedimentation assay of Cynomolgus Alpha Synuclein Pre-formed Fibrils: Biotinylated Protein (SPR-528)Thioflavin T assay of Cynomolgus Alpha Synuclein Pre-formed Fibrils: Biotinylated Protein (SPR-528)TEM of Cynomolgus Alpha Synuclein Pre-formed Fibrils: Biotinylated Protein (SPR-528)AFM of Cynomolgus Alpha Synuclein Pre-formed Fibrils: Biotinylated Protein (SPR-528)Biotinylation assay of Cynomolgus Alpha Synuclein Pre-formed Fibrils: Biotinylated Protein (SPR-528)
Product Name Alpha Synuclein Pre-formed Fibrils: Biotinylated
Description

Cynomolgus Recombinant Alpha Synuclein PFFs: Biotinylated (C-terminus)
Applications WB, SDS PAGE, In vitro Assay, In vivo Assay
Concentration 2 mg/mL
Conjugates C-terminal 15 amino acid tag: biotinylated
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
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

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
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

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
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

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
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications. 

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
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications. 

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
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

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*

*The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

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*
    • *The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

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

 

  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

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 Cynomolgus
Expression System E. coli
Amino Acid Sequence MDVFMKGLSKAKEGVVAAAEKTKQGVAEAAGKTKEGVLYVGSKTKEGVVHGVATVAEKTKEQVTNVGGAVVTGVTAVAQKTVEGAGSIAAATGFIKKDQLGKNEGAPQEGILQDMPVDPDNEAYEMPSEEGYQDYEPEAGLNDIFEAQKIEWHE
Purity >95%
Other Resources
Protein Length 155 amino acids
Protein Size 16.28 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 PBS 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
Cite This Product Cynomolgus Recombinant Alpha Synuclein Pre-formed Fibrils: Biotinylated (StressMarq Biosciences | Victoria, BC CANADA | Catalog# SPR-528)
Certificate of Analysis Protein certified > 95% pure via SDS-PAGE and nanodrop analysis. Low endotoxin (< 5 EU/mL) at 2 mg/mL.
Other Relevant Information For corresponding monomer see catalog# SPR-527 (unbiotinylated version not available yet but coming soon)

Biological Description

Alternative Names Alpha synuclein, Alpha-synuclein pre-formed fibril, Alpha synuclein protein PFF, Alpha synuclein PFF, 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, SYN protein, Parkinson disease familial 1 Protein
Research Areas Alzheimer's Disease, Neurodegeneration, Neuroscience, Parkinson's Disease, Synuclein, Tangles & Tau, Multiple System Atrophy
Cellular Localization Protein certified > 95% pure via SDS-PAGE and nanodrop analysis. Low endotoxin (< 5 EU/mL) at 2 & 5 mg/mL.
Swiss Prot P61142
Scientific Background A 15 amino acid tag on the C-terminal tail of alpha synuclein facilitates site-specific covalent biotinylation. Biotinylation of purified alpha synuclein allows for many biological applications, such as monitoring and detection using streptavidin-based conjugates. Stressmarq’s pre-formed fibrils are generated from biotinylated alpha synuclein monomers.
References 1.,Fairhead & Howarth. 2015. Site-specific biotinylation of purified proteins using BirA. Methods in Molecular Biology. DOI: 1007/978-1-4939-2272-7_12
2.,Hallacli et al. 2022. The Parkinson’s disease protein alpha synuclein is a modulator of Processing-bodies and mRNA stability. Cell. DOI: 10.1016/j.cell.2022.05.008
3.,Li et al. 2019. Naturally occurring antibodies isolated from PD patients inhibit synuclein seeding in vitro and recognize Lewy pathology. Acta neuropathologica. DOI: 10.1007/s00401-019-01974-5.

Product Images

<p>SDS-PAGE analysis of the starting monomers used for Cynomolgus Recombinant Alpha Synuclein Pre-Formed Fibrils: Biotinylated (C-Terminus) on a 12% Bis-Tris gel.</p>

SDS-PAGE analysis of the starting monomers used for Cynomolgus Recombinant Alpha Synuclein Pre-Formed Fibrils: Biotinylated (C-Terminus) on a 12% Bis-Tris gel.

<p>Representative TEM image of Cynomolgus Alpha Synuclein Pre-Formed Fibrils: Biotinylated (C-Terminus), 100nm scale. Negative stain transmission electron microscopy images of SPR-528 acquired at 80 Kv on carbon coated 400 mesh copper grids using phosphotungstic acid and uranyl acetate stain.</p>

Representative TEM image of Cynomolgus Alpha Synuclein Pre-Formed Fibrils: Biotinylated (C-Terminus), 100nm scale. Negative stain transmission electron microscopy images of SPR-528 acquired at 80 Kv on carbon coated 400 mesh copper grids using phosphotungstic acid and uranyl acetate stain.

<p>Sedimentation assay on Cynomolgus Alpha Synuclein Pre-Formed Fibrils: Biotinylated (C-Terminus). Samples were spun down at 15,000 x g, washed, and then spun down again. Fibril samples are prepared in denaturing conditions prior to running on the gel. SDS-PAGE analysis on a 12% Bis-Tris gel shows that the majority of the fibril is insoluble.</p>

Sedimentation assay on Cynomolgus Alpha Synuclein Pre-Formed Fibrils: Biotinylated (C-Terminus). Samples were spun down at 15,000 x g, washed, and then spun down again. Fibril samples are prepared in denaturing conditions prior to running on the gel. SDS-PAGE analysis on a 12% Bis-Tris gel shows that the majority of the fibril is insoluble.

<p>In vitro seeding activity of Cynomolgus Alpha Synuclein Pre-Formed Fibrils: Biotinylated (C-Terminus) in a Thioflavin T (ThT) assay. Cynomolgus Alpha Synuclein Pre-Formed Fibrils: Biotinylated (C-Terminus) (SPR-528) seed fibril formation of Cynomolgus Alpha Synuclein Monomers: C-Term Biotinylated (SPR-527) 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℃ in the presence of 25 uM ThT and read with an XPS Microplate Reader set at 450nmex/485nmem.</p>

In vitro seeding activity of Cynomolgus Alpha Synuclein Pre-Formed Fibrils: Biotinylated (C-Terminus) in a Thioflavin T (ThT) assay. Cynomolgus Alpha Synuclein Pre-Formed Fibrils: Biotinylated (C-Terminus) (SPR-528) seed fibril formation of Cynomolgus Alpha Synuclein Monomers: C-Term Biotinylated (SPR-527) 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℃ in the presence of 25 uM ThT and read with an XPS Microplate Reader set at 450nmex/485nmem.

<p>Representative TEM image of Cynomolgus Recombinant Alpha Synuclein Pre-Formed Fibrils: Biotinylated (C-Terminus), 500nm scale. Negative stain transmission electron microscopy images of SPR-528 acquired at 80 Kv on carbon coated 400 mesh copper grids using phosphotungstic acid and uranyl acetate stain.</p>

Representative TEM image of Cynomolgus Recombinant Alpha Synuclein Pre-Formed Fibrils: Biotinylated (C-Terminus), 500nm scale. Negative stain transmission electron microscopy images of SPR-528 acquired at 80 Kv on carbon coated 400 mesh copper grids using phosphotungstic acid and uranyl acetate stain.

<p>Atomic force microscopy analysis of 1 mg/mL SPR-528 diluted to 0.1 mg/mL with dH2O mounted on freshly cleaved mica, washed, dried and analyzed with tapping mode. Representative images are 10 x 10 µm x-y (left) and 5 x 5 µm x-y (right).</p>

Atomic force microscopy analysis of 1 mg/mL SPR-528 diluted to 0.1 mg/mL with dH2O mounted on freshly cleaved mica, washed, dried and analyzed with tapping mode. Representative images are 10 x 10 µm x-y (left) and 5 x 5 µm x-y (right).

<p>Biotinylation assay on purified Cynomolgus Alpha Synuclein Monomers: Biotinylated (C-Terminus). The assay is performed on 123 uM of monomers post-biotinylation (SPR-527), which is the starting material used to generate the Pre-Formed Fibrils (SPR-528), with 100 uM of biotin as a positive control & PBS pH 7.4 as a negative control. Absorbance is measured at 500nm, and results are converted to % of biotinylation.</p>

Biotinylation assay on purified Cynomolgus Alpha Synuclein Monomers: Biotinylated (C-Terminus). The assay is performed on 123 uM of monomers post-biotinylation (SPR-527), which is the starting material used to generate the Pre-Formed Fibrils (SPR-528), with 100 uM of biotin as a positive control & PBS pH 7.4 as a negative control. Absorbance is measured at 500nm, and results are converted to % of biotinylation.

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