Tau (K18) P301L Mutant Pre-formed Fibrils

Name: Tau (K18) P301L Mutant Pre-formed Fibrils
SKU: SPR-523
Availability: InStock

Human Recombinant Tau (K18) P301L Mutant Pre-Formed Fibrils (fibrillized without heparin)

Catalog No. SPR-523

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Expression System	E. coli
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SKU: SPR-523 Category: Proteins

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TEM of Tau (K18) P301L Mutant Pre-formed Fibrils (SPR-523)

Native-PAGE of Tau (K18) P301L Mutant Pre-formed Fibrils (SPR-523)

Thioflavin T assay of Tau (K18) P301L Mutant Pre-formed Fibrils (SPR-523)

AFM of Tau (K18) P301L Mutant Pre-formed Fibrils (SPR-523)

Toxicity of Tau (K18) P301L Mutant Pre-formed Fibrils (SPR-523)

Product Name Tau (K18) P301L Mutant Pre-formed Fibrils

Description

Human Recombinant Tau (K18) P301L Mutant Pre-Formed Fibrils (fibrillized without heparin)

Applications WB, SDS-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:

λ_ex = 493 nm

λ_em = 518 nm

ε_max = 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:

λ_ex = 652 nm

λ_em = 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:

λ_ex = 353 nm

λ_em = 432 nm

ε_max = 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:

λ_ex = 400 nm

λ_em = 420 nm

ε_max = 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:

λ_ex = 593 nm

λ_em = 618 nm

ε_max = 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:

λ_ex = 638 nm

λ_em = 658 nm

ε_max = 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:

λ_ex = 482 nm

λ_em = 677 nm

ε_max = 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:

λ_ex = 535 nm

λ_em = 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:

λ_ex = 494 nm

λ_em = 520 nm

ε_max = 7.3×10⁴

Φ_f = 0.92

τ_fl = 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:

λ_ex = 700 nm

λ_em = 719 nm

ε_max = 1.25×10⁵

Φ_f = 0.25

τ_fl = 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:

λ_ex = 680 nm

λ_em = 700 nm

ε_max = 1.25×10⁵

Φ_f = 0.30

τ_fl = 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:

λ_ex = 663 nm

λ_em = 684 nm

ε_max = 1.25×10⁵

Φ_f = 0.30

τ_fl = 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:

λ_ex = 629 nm

λ_em = 657 nm

ε_max = 1.3×10⁵

Φ_f = 0.64

τ_fl = 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:

λ_ex = 601 nm

λ_em = 627 nm

ε_max = 1.2×10⁵

Φ_f = 0.85

τ_fl = 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:

λ_ex = 563 nm

λ_em = 592 nm

ε_max = 1.2×10⁵

Φ_f = 0.9

τ_fl = 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:

λ_ex = 501 nm

λ_em = 523 nm

ε_max = 9.0×10⁴

Φ_f = 0.80

τ_fl = 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:

λ_ex = 390 nm

λ_em = 479 nm

ε_max = 2.4×10⁴

Φ_f = 0.90

τ_fl = 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:

λ_ex = 650 nm

λ_em = 660 nm

ε_max = 7.0×10⁵

Φ_f = 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

Biotin

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:

λ_ex = 565 nm

λ_em = 575 nm

ε_max = 2.0×10⁶

Φ_f = 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 MSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIINKKLDLSNVQSKCGSKDNIKHVLGGGSVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRE

Purity >95%

Other Resources

Protein Length 141 amino acids

Protein Size 15.2 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	10mM HEPES pH 7.4, 100mM NaCl
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 (K18) P301L Mutant Pre-formed Fibrils (StressMarq Biosciences \| Victoria, BC CANADA \| Catalog# SPR-523)
Certificate of Analysis	Protein certified > 95% pure via SDS-PAGE and A260/A280 ratio
Other Relevant Information	Monomer source is SPR-328

Biological Description

Alternative Names	Tau K18 P301L, K18-P301L Tau, Tau 4R repeat domain P301L, Tau MTBR P301L, Tau microtubule-binding region P301L, Tau repeat domain P301L (Tau-RD P301L), Tau K18 mutant P301L, Recombinant Tau K18 P301L fragment, Tau 4-repeat construct P301L, Tau RD(P301L), Tau K18 familial FTDP-17 mutant, Tau PFFs
Research Areas	Alzheimer's Disease, Neurodegeneration, Neuroscience, Tangles & Tau
Swiss Prot	P10636
Scientific Background	Tauopathies are a class of neurodegenerative diseases characterized by the pathological aggregation of tau protein. The K18 fragment, comprising the microtubule-binding repeat domains of tau, is frequently used in experimental models due to its propensity to form fibrillar aggregates. The P301L mutation, located within the repeat domain, has been demonstrated to increase the protein's propensity to aggregate and seed, making it a key variant in modeling tau pathology. Our Tau (K18) P301L Mutant Pre-formed Fibrils are fibrillized without heparin and have been demonstrated to be neurotoxic to primary mouse cortical neurons.
References	1.,Peeraer, E., Bottelbergs, A., Van Kolen, K., Stancu, I. C., Vasconcelos, B., Mahieu, M., Duytschaever, H., Ver Donck, L., Torremans, A., Sluydts, E., Van Acker, N., Kemp, J. A., Mercken, M., Brunden, K. R., Trojanowski, J. Q., Dewachter, I., Lee, V. M., & Moechars, D. (2015). Intracerebral injection of preformed synthetic tau fibrils initiates widespread tauopathy and neuronal loss in the brains of tau transgenic mice. Neurobiology of disease, 73, 83–95. https://doi.org/10.1016/j.nbd.2014.08.032 2.,Croft, C. L., Goodwin, M. S., Ryu, D. H., Lessard, C. B., Tejeda, G., Marrero, M., Vause, A. R., Paterno, G., Cruz, P. E., Lewis, J., Giasson, B. I., & Golde, T. E. (2021). Photodynamic studies reveal rapid formation and appreciable turnover of tau inclusions. Acta Neuropathologica, 141, 359–381. https://doi.org/10.1007/s00401-021-02264-9 3.,Zhang, X., Wang, J., Zhang, Z., & Ye, K. (2024). Tau in neurodegenerative diseases: molecular mechanisms, biomarkers, and therapeutic strategies. Translational Neurodegeneration, 13(40). https://doi.org/10.1186/s40035-024-00429-6

Product Images

<p>SDS-PAGE analysis of purified Tau (K18) P301L Mutant Monomers (Cat # SPR-328), on a 12% Bis-Tris gel. These monomers are used as the starting material for Cat # SPR-523.</p>

SDS-PAGE analysis of purified Tau (K18) P301L Mutant Monomers (Cat # SPR-328), on a 12% Bis-Tris gel. These monomers are used as the starting material for Cat # SPR-523.

<p>Representative TEM image of Tau (K18) P301L Mutant Pre-Formed Fibrils, 200nm scale. Negative stain transmission electron microscopy images of SPR-523 acquired at 80 Kv on carbon coated 400 mesh copper grids using phosphotungstic acid and uranyl acetate stain.</p>

Representative TEM image of Tau (K18) P301L Mutant Pre-Formed Fibrils, 200nm scale. Negative stain transmission electron microscopy images of SPR-523 acquired at 80 Kv on carbon coated 400 mesh copper grids using phosphotungstic acid and uranyl acetate stain.

<p>Native-PAGE of Tau (K18) P301L Mutant Monomers (SPR-328) & Pre-Formed Fibrils (SPR-523) on a 12% Tris-glycine gel (no SDS). Note: native proteins typically migrate much slower through a gel compared to reduced, SDS-treated MW standards.</p>

Native-PAGE of Tau (K18) P301L Mutant Monomers (SPR-328) & Pre-Formed Fibrils (SPR-523) on a 12% Tris-glycine gel (no SDS). Note: native proteins typically migrate much slower through a gel compared to reduced, SDS-treated MW standards.

<p>In vitro seeding activity of Tau (K18) P301L Mutant Pre-Formed Fibrils in a ThT assay. Tau (K18) P301L Mutant Pre-Formed Fibrils (SPR-523) seed fibril formation of Tau (K18) P301L Mutant Pre-Formed Fibrils (SPR-328) over 72 hours. Reactions (100uL) are shaken at 600 rpm in Greiner-Bio 96 Well Non-Binding Cell Culture Microplates, Black (Greiner-Bio Catalog #655900) at 37C in the presence of 25 uM ThT and read with an XPS Microplate Reader set at 450nm ex/485nm em.</p>

In vitro seeding activity of Tau (K18) P301L Mutant Pre-Formed Fibrils in a ThT assay. Tau (K18) P301L Mutant Pre-Formed Fibrils (SPR-523) seed fibril formation of Tau (K18) P301L Mutant Pre-Formed Fibrils (SPR-328) over 72 hours. Reactions (100uL) are shaken at 600 rpm in Greiner-Bio 96 Well Non-Binding Cell Culture Microplates, Black (Greiner-Bio Catalog #655900) at 37C in the presence of 25 uM ThT and read with an XPS Microplate Reader set at 450nm ex/485nm em.

<p>Representative TEM image of Tau (K18) P301L Mutant Pre-Formed Fibrils, 500nm scale. Negative stain transmission electron microscopy images of SPR-523 acquired at 80 Kv on carbon coated 400 mesh copper grids using phosphotungstic acid and uranyl acetate stain.</p>

Representative TEM image of Tau (K18) P301L Mutant Pre-Formed Fibrils, 500nm scale. Negative stain transmission electron microscopy images of SPR-523 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-523 diluted to 0.1 mg/mL with ultrapure H2O mounted on freshly cleaved mica, washed, dried and analyzed with tapping mode. Representative images are 5 x 5 µm x-y (left) and 2 x 2 µm x-y (right), both with a Z-range set at 11nm.</p>

Atomic force microscopy analysis of 1 mg/mL SPR-523 diluted to 0.1 mg/mL with ultrapure H2O mounted on freshly cleaved mica, washed, dried and analyzed with tapping mode. Representative images are 5 x 5 µm x-y (left) and 2 x 2 µm x-y (right), both with a Z-range set at 11nm.

<p>Tau (K18) P301L Mutant Pre-Formed Fibrils (average = 49.7%, SD = 9.6, n = 6) demonstrate increased toxicity to primary mouse cortical neurons compared to the Tau (K18) P301L Mutant Monomers (average = 97.9%, SD = 14.9, n = 11). The protein constructs and a control (10 µM Afatinib) were added to the neuron cultures at 12 DIV and toxicity assessed via a CCK-8 assay at 15 DIV.</p>

Tau (K18) P301L Mutant Pre-Formed Fibrils (average = 49.7%, SD = 9.6, n = 6) demonstrate increased toxicity to primary mouse cortical neurons compared to the Tau (K18) P301L Mutant Monomers (average = 97.9%, SD = 14.9, n = 11). The protein constructs and a control (10 µM Afatinib) were added to the neuron cultures at 12 DIV and toxicity assessed via a CCK-8 assay at 15 DIV.

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Tau (K18) P301L Mutant Pre-formed Fibrils

Streptavidin

Biotin

HRP (Horseradish peroxidase)

AP (Alkaline Phosphatase)

Properties

Biological Description

Product Images

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