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Thioflavin T

Thioflavin T is a fluorescent dye widely used in the study of amyloid fibrils and protein aggregattion.
It binds to beta-sheet rich structures, enabling visualization and quantification of these protein assemblies.
Thioflavin T research is crucial for understanding neurodegenerative diseases and other amyloid-related disorders.
PubCompare.ai enhances reproducibility and accuracy in Thioflavin T studies by providing easy access to protocols from literature, pre-prints, and patents, along with AI-driven comparisons to identify the best methods and products for your research.
Leveraging this platform can take your Thioflavin T studies to the next level and accelerate your discoveries.

Most cited protocols related to «Thioflavin T»

1
Quantifying Amyloid Fibrils with ThT
Cited 29 times
ThT (Sigma, product no. T3516) was dissolved in PBS buffer and was filtered through a 0.2 µm syringe filter. Then the concentration of thioflavin T was determined using an extinction coefficient of 36 mM−1 cm−1 at 412 nm [27 ,28 (link)].
For measurement of ThT fluorescence without proteins, various concentrations of ThT were prepared in PBS using serial dilution. Forty microlitres of ThT sample at each concentration was transferred to a black 384-well Nonbinding Surface microplate with clear bottom (Corning product no. 3655). The ThT fluorescence was measured at room temperature (approx. 24°C) using a Victor 3 V plate reader (Perkin Elmer) through the bottom of the plate with excitation filter of 450 nm and emission filter of 490 nm.
For measurement of ThT fluorescence in the presence of amyloid fibrils, various concentrations of sonicated Aβ40, Aβ42 and Ure2 fibrils were also prepared using serial dilution. Then 20 µl of ThT was mixed with 20 µl of sonicated fibrils to achieve desired concentrations of ThT and amyloid. Fluorescence measurements were performed as described above.
Xue C., Lin T.Y., Chang D, & Guo Z. (2017). Thioflavin T as an amyloid dye: fibril quantification, optimal concentration and effect on aggregation. Royal Society Open Science, 4(1), 160696.
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Publication 2017
Amyloid Fibrils Amyloid Proteins Buffers Extinction, Psychological Fluorescence MM 36 Proteins Syringes Technique, Dilution thioflavin T
2
Purification and Aggregation of α-Synuclein Fibrils
Cited 29 times

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Publication 2011
Amino Acids Amyloid Proteins Fluorometry Hyperostosis, Diffuse Idiopathic Skeletal Neurons Peptides thioflavin T
3
Characterization of α-Synuclein Aggregates
Cited 29 times
For fibril formation, soluble WT α-syn was incubated in buffer A (50 mM Tris-HCl, pH 7.5, 150 mM KCl) at 37 °C under continuous shaking in an Eppendorf Thermomixer set at 600 r.p.m. Assembly was monitored continuously in a Cary Eclipse spectrofluorimeter (Varian Inc., Palo Alto, CA, USA) in the presence of Thioflavin T (15 μM) in 1 × 1 cm cuvettes under agitation (100 r.p.m.) using a magnetic stir bar (6 × 3 mm) with excitation wavelength set at 440 and emissions wavelengths set at 440 and 480 nm, and an averaging time of 1 s.
For ribbon formation, WT α-syn was dialysed 16 h against 1,000 volume of buffer B (5 mM Tris-HCl pH 7.5) at 4 °C, then incubated at 37 °C under continuous shaking in an Eppendorf Thermomixer set at 600 r.p.m. Assembly was monitored by the measurement of the scattered light at 440 nm. Alternatively, the amount of protein remaining in the supernatant after sedimentation at 35,000 g was determined by measurement of the absorbance at 280 nm in a Hewlett Packard 8453 diode array spectrophotometer.
The nature of the oligomeric species was assessed using a Jeol 1400 (Jeol Ltd.)TEM following adsorption of the samples onto carbon-coated 200-mesh grids and negative staining with 1% uranyl acetate. The images were recorded with a Gatan Orius CCD camera (Gatan).
The ability of α-syn assemblies to bind Congo red was assessed as follows: α-syn fibrils and ribbons were incubated for 1 h with 100 μM Congo Red (Sigma-Aldrich, St Louis, MO, USA) in 20 mM Tris buffer (pH 7.5). The polymers were then sedimented at 20 °C in a TL100 Tabletop Beckman ultracentrifuge (Beckman Instruments, Inc., Fullerton, CA, USA) at 25,000 g for 30 min. The pellets were washed four times using an equal volume of water. Following resuspension of the pellets an aliquot was placed on a glass coverslip and imaged immediately or allowed to dry. Samples were viewed in bright field and cross-polarized light by polarization microscopy using a Leica (MZ12.5) microscope equipped with cross-polarizers (Leica Microsystems, Ltd., Heerbrugg, Switzerland).
Bousset L., Pieri L., Ruiz-Arlandis G., Gath J., Jensen P.H., Habenstein B., Madiona K., Olieric V., Böckmann A., Meier B.H, & Melki R. (2013). Structural and functional characterization of two alpha-synuclein strains. Nature Communications, 4, 2575.
Publication 2013
Adsorption Buffers Carbon Light Light Microscopy Microscopy Pellets, Drug Polymers Proteins thioflavin T Tromethamine uranyl acetate
4
Neuropathological and Genetic Characterization of Alzheimer's Disease Subtypes
Cited 15 times
Gross and macroscopic neuropathologic assessment was performed by standardized procedures. Formalin-fixed, paraffin-embedded tissue samples from the primary motor and visual cortices, inferior parietal lobule, mid-frontal gyrus, superior temporal gyrus, amygdala, and posterior hippocampus were cut at 5 µm thickness, mounted on glass slides and stained with H&E. Thioflavin-S fluorescent microscopy was performed to evaluate SP (10× objective) and NFT densities (40× objective). Primitive, neuritic, and cored type plaques were included in the SP counts and were truncated at 50, which is twice the number required to meet Khachaturian’s criteria for AD diagnosis [20 (link)]. NFT distribution was assessed to determine Braak stage. Intracellular and extracellular NFT counts from two hippocampal regions (CA1 and subiculum) and three association cortices (inferior parietal, mid-frontal, and superior temporal) were used to determine AD subtypes. Additional sections previously underwent immunohistochemical staining and were processed using the DAKO Autostainer (DAKO Auto Machine Corporation, Carpinteria, CA, USA) with DAKO Envision+ HRP System. The posterior hippocampus was stained with an antibody that detects the 25 kDa C-terminal fragment of TDP-43 (a generous gift from Leonard Petrucelli, Mayo Clinic, FL, USA). Assessment of TDP-43 pathology was performed as previously described [2 (link)]. Cerebrovascular disease was assessed using a simple scheme proposed by Jellinger and Attems [18 ], as previously reported. Lewy body disease (LBD) pathology was assessed by use of immunohistochemistry [34 (link)], as previously described.
For genotyping, genomic DNA was extracted from frozen brain by standard procedures. Genotyping for MAPT H1/H2 (SNP rs1052554 A/G, A = H1, G = H2), APOE alleles (SNP rs429358 C/T and rs7412 C/T), GRN (SNP rs5848 C/T), and TMEM106B (SNP rs1990622 C/T) was performed using a Taqman SNP genotyping assay (Applied Biosystems, Carlsbad, CA, USA). Genotype calls were obtained with SDS v2.2 software (Applied Biosystems). Although there is overlap in the HpScl-AD group between previous Genetic studies [9 (link), 32 (link)], Genetic information on the HpScl group has not been previously reported. Genotyping availability can be found in Supplementary methods.
Clinical reports were reviewed blind to pathologic diagnosis to collect education, family history, age of onset, disease duration, and Mini Mental State Examination (MMSE) scores. Family history was noted as positive if at least one first-degree relative had dementia. Age of onset was recorded as the age of initial cognitive abnormalities, as opposed to age of diagnosis. Disease duration was ascertained as the number of years elapsed between age of death and age of onset. Longitudinal decline was calculated as a slope of three or more MMSE scores, where the MMSE score was the dependent variable and elapsed years between testing and death were the independent variable. Antemortem clinical diagnosis of probable AD, possible AD and mild cognitive impairment was considered an amnestic diagnosis. Availability of clinical information can be found in Supplementary methods.
Murray M.E., Cannon A., Graff-Radford N.R., Liesinger A.M., Rutherford N.J., Ross O.A., Duara R., Carrasquillo M.M., Rademakers R, & Dickson D.W. (2014). Differential clinicopathologic and Genetic features of late-onset amnestic dementias. Acta neuropathologica, 128(3), 411-421.
Publication 2014
Alleles Amygdaloid Body Antemortem Diagnosis Apolipoproteins E Biological Assay Blindness Brain Cerebrovascular Disorders Cognition Cognitive Impairments, Mild Congenital Abnormality Cortex, Cerebral Dementia Diagnosis Formalin Freezing Genome Immunoglobulins Immunohistochemistry Lewy Body Disease MAPT protein, human Microscopy Mini Mental State Examination Neuritis Paraffin Embedding Parietal Lobule protein TDP-43, human Protoplasm Seahorses Senile Plaques Subiculum Superior Temporal Gyrus thioflavin S Visual Cortex
5
Optimized RT-QuIC Assay for Prion Detection
Cited 14 times
RT-QuIC reactions were performed as previously described [14 (link)]. Reaction mix was composed of 10 mM phosphate buffer (pH 7.4), 300 or 130 mM NaCl, 0.1 mg/ml rPrPSen, 10 μM thioflavin T (ThT), 1 mM ethylenediaminetetraacetic acid tetrasodium salt (EDTA), and 0.002% or 0.001% SDS. NaCl and SDS concentrations were varied where indicated. Aliquots of the reaction mix (98 μL) were loaded into each well of a black 96-well plate with a clear bottom (Nunc) and seeded with 2 μL of indicated BH dilutions. The plate was then sealed with a plate sealer film (Nalgene Nunc International) and incubated at 42°C in a BMG FLUOstar Omega plate reader with cycles of 1 min shaking (700 rpm double orbital) and 1 min rest throughout the indicated incubation time. ThT fluorescence measurements (450 +/-10 nm excitation and 480 +/-10 nm emission; bottom read) were taken every 45 min.
To compensate for minor differences in baselines between fluorescent plate readers and across multiple experiments, data sets were normalized to a percentage of the maximal fluorescence response (260,000 rfu) of the plate readers after subtraction of the baseline, as described [34 (link)], and plotted versus reaction time. Reactions were classified as RT-QuIC positive base on criteria similar to those previously described for RT-QuIC analyses of brain specimens [14 (link),34 (link)].
Orrú C.D., Groveman B.R., Raymond L.D., Hughson A.G., Nonno R., Zou W., Ghetti B., Gambetti P, & Caughey B. (2015). Bank Vole Prion Protein As an Apparently Universal Substrate for RT-QuIC-Based Detection and Discrimination of Prion Strains. PLoS Pathogens, 11(6), e1004983.
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Publication 2015
Brain Buffers Edetic Acid Fluorescence Phosphates Salts Sodium Chloride Technique, Dilution thioflavin T

Most recents protocols related to «Thioflavin T»

1
Thioflavin T Staining of C. albicans Infection
iBMDM (2*105 cells/well) were seeded in a 24-well plate overnight and then infected with C. albicans, or treated with LPS or thapsigargin for indicated timepoints. Thioflavin T (Cayman Chemical, 5 μM) was added 2 hours prior to endpoint. Cells were scraped into ice cold PBS and Thioflavin T intensity was measured on a BD LSRFortessa X-20 flow cytometer.
McFadden M.J., Reynolds M.B., Michmerhuizen B.C., Ólafsson E.B., Anderson F.M., Schultz T.L., O’Riordan M.X, & O’Meara T.R. (2024). Non-canonical activation of IRE1α during Candida albicans infection enhances macrophage fungicidal activity. bioRxiv.
Publication Preprint 2024
2
Quantifying Neurofibrillary Tangles via Thioflavin T
Thioflavin T staining method was employed to quantify the levels of neurofibrillary tangles (NFTs). Tissue sections were brought to room temperature and washed three times for 5 min each in tbs (A510025, Sangon Biotech). Then, the tissue sections were immersed in a light-protected solution of 0.0125% Thioflavin T in ethanol (HY-D0972, MedChemExpress) for 8 min. Subsequently, the tissue sections were washed three times for 5 min each in 50% ethanol, followed by one wash in TBS for 5 min. After drying the tissue sections in a light-protected manner, glycerin was used for sealing. The staining results were observed and photographed under a fluorescence microscope (Olympus, Tokyo, Japan).
Xie X., Zhang X., Li S, & Du W. (2024). Involvement of Fgf2-mediated tau protein phosphorylation in cognitive deficits induced by sevoflurane in aged rats. Molecular Medicine, 30, 39.
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Publication 2024
3
Thioflavin T Assay for Protein Aggregation
Proteins were dissolved in phosphate buffered saline, pH 7.2 (Sigma‐Aldrich). Thioflavin T was prepared fresh for each assay at a concentration of 400 μM in DMSO, then diluted to 20 μM in PBS/Tris. Twenty‐five microliters of protein and ThT in PBS/Tris were combined in a 96‐well plate (Catalog 3596, Costar, Fisher Scientific) and incubated for 15 min at room temperature in the dark. Fluorescence was measured using a plate reader (Spark 10 M, Tecan) with an excitation at 440 nm, emission was collected at 486 nm. Assays performed using Tris rather than PBS were those measuring CLC due to solubility issues, which was resuspended in 20 mM Tris buffer pH 7.5. For these assays, Thioflavin T was also diluted in this buffer. All proteins and controls were assayed in triplicate.
Sanchez‐Martinez S., Nguyen K., Biswas S., Nicholson V., Romanyuk A.V., Ramirez J., Kc S., Akter A., Childs C., Meese E.K., Usher E.T., Ginell G.M., Yu F., Gollub E., Malferrari M., Francia F., Venturoli G., Martin E.W., Caporaletti F., Giubertoni G., Woutersen S., Sukenik S., Woolfson D.N., Holehouse A.S, & Boothby T.C. (2024). Labile assembly of a tardigrade protein induces biostasis. Protein Science : A Publication of the Protein Society, 33(4), e4941.
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Publication 2024
4
Hydrogel-based Thioflavin T Assay
200 μL hydrogel was heated to a transparent liquid and mixed with 2 μL Thioflavin T (ThT) solution (1 mmol). Then, the sample was cooled to room temperature to form hydrogels. The fluorescence of the sample was observed at 365 nm by a dark-box ultraviolet analyzer (JY02S, Beijing Junyi, China), and detected by a Cary Eclipse fluorescence spectrophotometer (Agilent, USA) with excitation/emission wavelengths at 450/485 nm.
Li W., Wen Y., Wang K., Ding Z., Wang L., Chen Q., Xie L., Xu H, & Zhao H. (2024). Developing a machine learning model for accurate nucleoside hydrogels prediction based on descriptors. Nature Communications, 15, 2603.
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Publication 2024
5
Visualizing Fmoc-FF Fiber Scaffolds
Not available on PMC !
Thioflavin T (ThT) dye (Sigma Aldrich) was used to visualise Fmoc-FF fibres within the hybrid scaffolds with laser scanning confocal microscopy. 200 µL samples were bioprinted into 8 chamber microscopy slides (Lab-Tek) and then stained overnight with 25 µM ThT and washed 6 times with TBS before being imaged. Zeiss LSM 780 microscope (Zeiss, Germany) was used to image the samples at an excitation wavelength of 450 nm and an emission wavelength of 485 nm and a 40x water immersion lens (NA = 1.1) at room temperature.
Field E.H., Ratcliffe J., Johnson C., Binger K.J., & Reynolds N.P. (2024). Self-healing, biocompatible bioinks from self-assembled peptide and alginate hybrid hydrogels.
Publication 2024

Top products related to «Thioflavin T»

1
Thioflavin t by Merck Group
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Thioflavin T is a fluorescent dye used in the detection and quantification of amyloid fibrils. It exhibits enhanced fluorescence upon binding to these protein aggregates. The dye is commonly utilized in various research applications, including the study of protein misfolding and amyloidosis.
2
Thioflavin t tht by Merck Group
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Thioflavin T (ThT) is a fluorescent dye used as a research tool. It has the ability to bind to amyloid fibrils, a type of protein aggregation, and emit fluorescence upon binding. The core function of ThT is to serve as a detection and quantification method for amyloid formation in various biological samples.
3
Fluostar omega by BMG Labtech
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The FLUOstar Omega is a multimode microplate reader designed for a variety of fluorescence, luminescence, and absorbance-based applications. It offers high-performance detection capabilities and supports a wide range of microplate formats.
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Congo red by Merck Group
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Congo red is a synthetic dye used as a laboratory reagent. It is a dark red crystalline powder that is soluble in water and certain organic solvents. Congo red is commonly used as an indicator in various analytical and diagnostic applications, particularly in the identification of amyloid proteins.
5
Dimethyl sulfoxide (dmso) by Merck Group
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DMSO is a versatile organic solvent commonly used in laboratory settings. It has a high boiling point, low viscosity, and the ability to dissolve a wide range of polar and non-polar compounds. DMSO's core function is as a solvent, allowing for the effective dissolution and handling of various chemical substances during research and experimentation.
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The FLUOstar OPTIMA is a multi-mode microplate reader designed for a wide range of applications in life science research and drug discovery. It provides precise and accurate measurements of fluorescence, luminescence, and absorbance in microplates.
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Synergy h1 by Agilent Technologies
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The Synergy H1 is a multi-mode microplate reader designed for a variety of applications. It is capable of absorbance, fluorescence, and luminescence detection.
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Cary eclipse fluorescence spectrophotometer by Agilent Technologies
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The Cary Eclipse Fluorescence Spectrophotometer is a laboratory instrument designed to measure the fluorescence properties of samples. It is capable of performing excitation and emission scans, as well as time-based measurements. The instrument uses a xenon flash lamp as the light source and provides high-sensitivity detection and rapid scanning capabilities.
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Fluostar omega plate reader by BMG Labtech
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The FLUOstar Omega is a multimode microplate reader that can perform absorbance, fluorescence, and luminescence measurements. It offers high-quality optics and a monochromator-based excitation and emission system to provide flexible wavelength selection. The FLUOstar Omega is designed for a wide range of applications in life science research and drug discovery.

More about "Thioflavin T"

Thioflavin T, also known as ThT, is a widely used fluorescent dye in the study of amyloid fibrils and protein aggregation.
This compound has the unique ability to bind to beta-sheet rich structures, enabling researchers to visualize and quantify these protein assemblies.
This is crucial for understanding neurodegenerative diseases, such as Alzheimer's and Parkinson's, as well as other amyloid-related disorders.
The FLUOstar Omega, FLUOstar OPTIMA, and Synergy H1 are plate readers commonly used in Thioflavin T studies, as they allow for the accurate measurement and quantification of fluorescence.
Additionally, Congo red is another dye that is often used in conjunction with Thioflavin T to study amyloid structures.
Preparing Thioflavin T solutions can be done using DMSO, a commonly used solvent.
Bovine serum albumin (BSA) is also sometimes used as a stabilizing agent in Thioflavin T assays.
The Cary Eclipse Fluorescence Spectrophotometer is another instrument that can be utilized to analyze Thioflavin T-bound protein aggregates.
PubCompare.ai is an AI-driven platform that enhances the reproducibility and accuracy of Thioflavin T studies.
By providing easy access to protocols from literature, pre-prints, and patents, along with AI-driven comparisons to identify the best methods and products, PubCompare.ai can help researchers take their Thioflavin T studies to the next level and accelerate their discoveries.
Leveraging this platform can be a game-changer in your amyloid research endeavors.