Tau and Alpha Synuclein Co-Polymer Fibrils

Human Recombinant Tau-352 (fetal 0N3R) and Human Recombinant Alpha Synuclein Co-Polymer Fibrils

Catalog No. SPR-494

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Expression System E. coli
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Immuno-TEM of Human Tau and Alpha Synuclein co-polymer fibrils Protein (SPR-494)SDS-Page of Human Tau and Alpha Synuclein co-polymer fibrils Protein (SPR-494)Thioflavin T assay of Human Tau and Alpha Synuclein co-polymer fibrils Protein (SPR-494)
Product Name Tau and Alpha Synuclein Co-Polymer Fibrils
Description

Human Recombinant Tau-352 (fetal 0N3R) and Human Recombinant Alpha Synuclein Co-Polymer Fibrils

Applications WB, SDS PAGE, In vitro assay
Concentration Total Protein Concentration: 2mg/mL (1mg/ml of tau and 1mg/ml of aSyn)
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 Tau:
MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKAEEAGIGDTPSLEDEAAGHVTQARMVSKSKDGTGSDDKKAKGADGKTKIATPRGAAPPG
QKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPTREPKKVAVVRTPPKSPSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGG
KVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL

Asyn:
MDVFMKGLSKAKEGVVAAAEKTKQGVAEAAGKTKEGVLYVGSKTKEGVVHGVATVAEKTKEQVTNVGGAVVTGVTAVAQKTVEGAGSIAAATGFVKKDQLGKNEEGAP
QEGILEDMPVDPDNEAYEMPSEEGYQDYEPEA
Purity >95%
Other Resources Sonication Protocol
Protein Length Full Length (Tau 0N3R: 1-352 aa, Asyn: 1-140 aa)
Protein Size 0N3R: 36.76 kDa, ASYN: 14.46 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 1X 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 Human Recombinant Tau and Alpha Synuclein Co-Polymer Fibrils (StressMarq Biosciences Inc., Victoria BC CANADA, Catalog # SPR-494)
Certificate of Analysis Protein certified >95% pure on SDS-PAGE & Nanodrop analysis. Low endotoxin <5 EU/mL @ 2mg/mL.
Other Relevant Information For corresponding monomers, see catalog# SPR-321 and SPR-490

Biological Description

Alternative Names MAPT, Fetal Tau, 0N3R, neurofibrillary tangle protein, paired-helical filament, PHFs, SNCA, NACP, PARK1, asyn, alpha-synuclein, pre-formed fibril, PFFs, mixed fibrils
Research Areas Alzheimer's Disease, Neurodegeneration, Neuroscience, Parkinson's Disease, Synuclein, Tangles & Tau, Multiple System Atrophy
Swiss Prot P10636-2 and P37840-1
Scientific Background Brain-specific tau isoforms vary in the number of N-terminal inserts and C- terminal repeat domains due to alternative splicing of exons; only the shortest isoform of tau, 0N3R, is expressed in the fetal brain during neurogenesis (1). Tau and alpha-synuclein polymerize into amyloid fibrils to form filamentous inclusions in neurodegenerative diseases such as Alzheimer’s and Parkinson’s disease. Tau has been shown to interact with alpha-synuclein in vitro (2), with synergistic cross-seeding between tau and alpha-synuclein resulting in polymerization of each other into fibrillary amyloid lesions in neuronal cultures and in vivo (3,4). Recombinant tau and alpha-synuclein co-polymer fibrils have demonstrated a more widespread transmission of induced pathology in a rodent model of tauopathies compared to pure Tau or alpha-synuclein fibrils alone (5). These co-polymer fibrils have also shown enhanced alpha-synuclein aggregation in vitro, and more severe alpha-synuclein pathology and Parkinson’s disease-like symptoms in mice (6).
References 1. Goedert et al. Multiple Isoforms of Human Microtubule-associated Protein Tau: Sequences and Localization in Neurofibrilary Tangles of Alzheimer’s Disease. Neuron. 1989;3(4):519-526.
2. Jensen et al. α-synuclein Binds to Tau and Stimulates the Protein Kinase A-catalyzed Tau Phosphorylation of Serine Residues 262 and 356. 1999. JBC. 274(36): 25481-25489. DOI:https://doi.org/10.1074/jbc.274.36.25481
3. Giasson et al. Initiation and Synergistic Fibrillization of Tau and Alpha-Synuclein. Science. 2003; 300: 636-40. DOI: 10.1126/science.1082324
4. Guo et al. Distinct α-synuclein Strains Differentially Promote Tau Inclusions in Neurons. 2013. Cell. 154(1) doi:10.1016/j.cell.2013.05.057.
5. Williams et al. Differential Cross-seeding Properties of Tau and α-synuclein in Mouse Models of Tauopathy and Synucleinopathy. Brain Communications. 2020; 2(2):fcaa090. doi:10.1093/braincomms/fcaa090
6. Pan et al. Tau Accelerates α-synuclein Aggregation and Spreading in Parkinson’s Disease. 2022. Brain. Doi: 10.1093/brain/awac171

Product Images

<p>TEM of Fetal Tau 0N3R & Alpha Synuclein Co-Polymer Fibrils (SPR-494). Negative stain transmission electron microscopy images of SPR-494 acquired at 80 Kv on carbon coated 400 mesh copper grids using phosphotungstic acid and uranyl acetate stain.</p>

TEM of Fetal Tau 0N3R & Alpha Synuclein Co-Polymer Fibrils (SPR-494). Negative stain transmission electron microscopy images of SPR-494 acquired at 80 Kv on carbon coated 400 mesh copper grids using phosphotungstic acid and uranyl acetate stain.

<p>Bis-Tris SDS-PAGE (12%) of Fetal Tau 0N3R & Alpha Synuclein Co-Polymer Fibrils. MW ladder = Precision Plus Protein All Blue pre-stained standards.</p>

Bis-Tris SDS-PAGE (12%) of Fetal Tau 0N3R & Alpha Synuclein Co-Polymer Fibrils. MW ladder = Precision Plus Protein All Blue pre-stained standards.

<p>In vitro Seeding activity of Fetal Tau 0N3R & Alpha Synuclein Co-Polymer Fibrils in ThT Assay. Tau 0N3R and Alpha Synuclein co-polymer fibrils seed fibril formation of both Alpha Synuclein monomers and of a mixture of Alpha Synuclein and Tau 0N3R monomers 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°C and read with an XPS Microplate Reader set at 450nmex/485nmem.</p>

In vitro Seeding activity of Fetal Tau 0N3R & Alpha Synuclein Co-Polymer Fibrils in ThT Assay. Tau 0N3R and Alpha Synuclein co-polymer fibrils seed fibril formation of both Alpha Synuclein monomers and of a mixture of Alpha Synuclein and Tau 0N3R monomers 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°C and read with an XPS Microplate Reader set at 450nmex/485nmem.

<p>Immuno-TEM of Tau & Alpha Synuclein Co-Polymer Fibrils. Fibrils absorbed onto carbon- coated copper grids. Grids were blocked with 1% BSA and 0.1% Tween-20 in PBS, incubated sequentially with primary antibodies at 20 µg/mL in blocking buffer, washed, then incubated with secondary antibodies at 20 µg/mL in blocking buffer, washed and stained with 2% uranyl acetate as a negative stain. 6nm (asyn) and 12nm (tau) signals only appear together in the same fibril strand for the co-polymer fibril samples, and no antibody cross reactivity is observed in control alpha-synuclein or tau fibrils. Primary antibodies: Mouse anti-human asyn monoclonal (StressMarq SMC-532) and rabbit anti-human tau polyclonal (StressMarq SPC-802); Secondary antibodies: 6nm Colloidal Gold-conjugated Goat Anti-Mouse (Jackson ImmunoResearch 115-195- 146) and 12nm Colloidal Gold-conjugated Goat Anti-Rabbit (Jackson ImmunoResearch 111-205-144).</p>

Immuno-TEM of Tau & Alpha Synuclein Co-Polymer Fibrils. Fibrils absorbed onto carbon- coated copper grids. Grids were blocked with 1% BSA and 0.1% Tween-20 in PBS, incubated sequentially with primary antibodies at 20 µg/mL in blocking buffer, washed, then incubated with secondary antibodies at 20 µg/mL in blocking buffer, washed and stained with 2% uranyl acetate as a negative stain. 6nm (asyn) and 12nm (tau) signals only appear together in the same fibril strand for the co-polymer fibril samples, and no antibody cross reactivity is observed in control alpha-synuclein or tau fibrils. Primary antibodies: Mouse anti-human asyn monoclonal (StressMarq SMC-532) and rabbit anti-human tau polyclonal (StressMarq SPC-802); Secondary antibodies: 6nm Colloidal Gold-conjugated Goat Anti-Mouse (Jackson ImmunoResearch 115-195- 146) and 12nm Colloidal Gold-conjugated Goat Anti-Rabbit (Jackson ImmunoResearch 111-205-144).

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