Tau-441 (2N4R) Wild-Type Oligomers

Human Recombinant Tau-441 (2N4R) Wild-Type Oligomers (Baculovirus/Sf9)

Catalog No. SPR-497

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Expression System Baculovirus (Sf9)
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Native-PAGE of Tau-441 (2N4R) Wild-Type Oligomers (SPR-497)Toxicity of Tau-441 (2N4R) Wild-Type Oligomers (SPR-497)SDS-Page of Tau-441 (2N4R) Wild-Type Oligomers (SPR-497)Dot Blot of Tau-441 (2N4R) Wild-Type Oligomers (SPR-497)
Product Name Tau-441 (2N4R) Wild-Type Oligomers
Description

Human Recombinant Tau-441 (2N4R) Wild-Type Oligomers (Baculovirus/Sf9)
Applications WB, SDS-PAGE, 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 Baculovirus (Sf9)
Amino Acid Sequence MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKESPLQTPTEDGSEEPGSETSDAKSTPTAEDVTAPLVDEGAPGKQAAAQPHTEIPEGTTAEEAGIGDTPSLEDEAAGHVTQARMVSKSKDGTGSDDKKAKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPTREPKKVAVVRTPPKSPSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIINKKLDLSNVQSKCGSKDNIKHVPGGGSVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL
Purity >95%
Other Resources
Protein Length Full Length (1-441 aa)
Protein Size 45.84 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 Human Recombinant Tau-441 (2N4R) Wild-Type Oligomers (StressMarq Biosciences Inc., Victoria BC CANADA, Catalog # SPR-497)
Certificate of Analysis Certified >95% pure using SDS-PAGE and A260/A280 analysis. Low endotoxin <5 EU/mL @ 2mg/mL.
Other Relevant Information Monomer source is catalog# SPR-496. For corresponding PFFs, see catalog# SPR-498.

Biological Description

Alternative Names MAPT, intracellular neurofibrillary tangles, NFTs, paired helical filaments, PHFs, 2N4R
Research Areas Alzheimer's Disease, Neurodegeneration, Neuroscience, Tangles & Tau
Swiss Prot P10636-8
Scientific Background Tau (tubulin-associated unit) is normally located in the axons of neurons where it stabilizes microtubules. Tauopathies such as Alzheimer’s Disease (AD) are characterized by neurofibrillary tangles containing hyper-phosphorylated tau fibrils (1) with consensus that tau oligomers are the most toxic species initiating neurodegeneration (2). Hyper-phosphorylated tau can be generated via expression in the Sf9/Baculovirus system, with up to 20 sites confirmed by mass spectrometry and western blots with phospho-specific antibodies (3). Our Sf9/Baculovirus-expressed Tau 2N4R oligomers are formed without the use of heparin or another anionic scaffold.
References 1.,Iqbal K., Liu F., and Gong C.X. 2016. Tau and neurodegenerative disease: The story so far. Nat. Rev. Neurol. DOI: 10.1038/nrneurol.2015.225
2.,Puangmalai, N., Bhatt, N., Montalbano, M. et al. 2020. Internalization mechanisms of brain-derived tau oligomers from patients with Alzheimer’s disease, progressive supranuclear palsy and dementia with Lewy bodies. Cell Death Dis. DOI: 10.1038/s41419-020-2503-3
3.,Tepper et al. 2014. Oligomer Formation of Tau Protein Hyperphophorylated in Cells. The Journal of Biological Chemistry, DOI: 10.1074/jbc.M114.611368

Product Images

<p>Atomic force microscopy analysis of 2 mg/mL SPR-497 diluted to 0.2 mg/mL with dH2O mounted on freshly cleaved mica, washed, dried and analyzed with tapping mode. Representative images are 2 x 2 µm x-y (left) and 5 x 5 µm x-y (right), both with a Z-range set at 4nm.</p>

Atomic force microscopy analysis of 2 mg/mL SPR-497 diluted to 0.2 mg/mL with dH2O mounted on freshly cleaved mica, washed, dried and analyzed with tapping mode. Representative images are 2 x 2 µm x-y (left) and 5 x 5 µm x-y (right), both with a Z-range set at 4nm.

<p>Post-oligomerization molecular weight (MW) shift of SPR-497 oligomers observed on a (A) 12% Tris-Glycine native-PAGE and by (B) size-exclusion chromatography of SPR-497. By peak area, at least 80% of SPR-497 is oligomeric. SEC was performed on Superdex 200 10/300 GL Increase column in phosphate buffer pH 7.4. Note: Monomeric Tau 2N4R is an intrinsically disordered 45 kDa protein. Due to its extended confirmation in solution, migration of free monomeric Tau 2N4R is similar to that of a globular 220 kDa protein on Native PAGE and SEC.</p>

Post-oligomerization molecular weight (MW) shift of SPR-497 oligomers observed on a (A) 12% Tris-Glycine native-PAGE and by (B) size-exclusion chromatography of SPR-497. By peak area, at least 80% of SPR-497 is oligomeric. SEC was performed on Superdex 200 10/300 GL Increase column in phosphate buffer pH 7.4. Note: Monomeric Tau 2N4R is an intrinsically disordered 45 kDa protein. Due to its extended confirmation in solution, migration of free monomeric Tau 2N4R is similar to that of a globular 220 kDa protein on Native PAGE and SEC.

<p>Tau 2N4R oligomers (catalog # SPR-497) and fibrils (catalog # SPR-498) show a dose-dependent toxicity to primary rat hippocampal neurons. Survival of rat primary hippocampal neurons 14 days after treatment with different concentrations of (A) monomers, (B) oligomers or (C) fibrils quantified by MAP2 positive neurons and expressed as a percentage of control. Fibrils were initially sonicated in a Bioruptor. Test conditions were run in the same plate as untreated control, which consisted of buffer without Tau protein. Data expressed as mean +/- s.e.m. (n=6). A global analysis of the data was performed using a one-way ANOVA followed by Dunnett’s test; * p < 0.05, ** p < 0.01, *** p < 0.0002, **** p < 0.0001; stats vs control. </p>

Tau 2N4R oligomers (catalog # SPR-497) and fibrils (catalog # SPR-498) show a dose-dependent toxicity to primary rat hippocampal neurons. Survival of rat primary hippocampal neurons 14 days after treatment with different concentrations of (A) monomers, (B) oligomers or (C) fibrils quantified by MAP2 positive neurons and expressed as a percentage of control. Fibrils were initially sonicated in a Bioruptor. Test conditions were run in the same plate as untreated control, which consisted of buffer without Tau protein. Data expressed as mean +/- s.e.m. (n=6). A global analysis of the data was performed using a one-way ANOVA followed by Dunnett’s test; * p < 0.05, ** p < 0.01, *** p < 0.0002, **** p < 0.0001; stats vs control.

<p>SDS-PAGE analysis of denatured hTau 2N4R wildtype oligomers on a 12% Tris-Glycine gel.</p>

SDS-PAGE analysis of denatured hTau 2N4R wildtype oligomers on a 12% Tris-Glycine gel.

<p>Dot Blot of purified hTau 2N4R constructs (SPR-480, SPR-496, SPR-497) using Stressmarq’s SPC-801 (anti-Tau) and SMC-601 (anti-p202/205) comparing phosphorylation in E.coli-expressed and baculovirus/sf9-expressed material. Protein was blotted on nitrocellulose, incubated with 1:1000 primary antibodies and/or 1:4000 secondary antibodies. Secondary control is goat-anti rabbit:HRP. Exposed 1 second.</p>

Dot Blot of purified hTau 2N4R constructs (SPR-480, SPR-496, SPR-497) using Stressmarq’s SPC-801 (anti-Tau) and SMC-601 (anti-p202/205) comparing phosphorylation in E.coli-expressed and baculovirus/sf9-expressed material. Protein was blotted on nitrocellulose, incubated with 1:1000 primary antibodies and/or 1:4000 secondary antibodies. Secondary control is goat-anti rabbit:HRP. Exposed 1 second.

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