Aggregation kinetics were measured in Synergy H1 microplate reader (BioTek) in black, flat-bottom, 96-well plates (Nunc). Tau or tau4R variants (10 μM) were pre-incubated in the presence or absence of indicated chaperones for 10 min at 37°C. All proteins in the assay were buffer exchanged into the assay buffer (50 mM HEPES pH 7.4, 50 mM KCl, and 2 mM DTT). ThT (Sigma) at a final concentration of 10 μM was added, and the aggregation was induced by the addition of 2.5 μM freshly prepared heparin salt solution (Sigma). Aggregation reactions were run at 37°C with continuous shaking (567 rpm) and monitored by ThT fluorescence (excitation = 440 nm, emission = 485 nm, bandwidth), using an area scan mode with a 3 × 3 matrix for each well. Black, flat-bottom, 96-well plates (Nunc) sealed with optical adhesive film (Applied Biosystems) were used. For data processing, baseline curves at same conditions but without heparin were subtracted from the data. Samples were run in triplicate, and the experiments were repeated at least four times with similar results.
Heparin salt solution
Manufactured by Merck Group
Heparin salt solution is a laboratory reagent used in various biochemical and analytical applications. It is a sterile, aqueous solution containing heparin, a naturally occurring anticoagulant. The solution is designed to maintain the stability and activity of heparin for use in research and laboratory settings.
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Lab products found in correlation
2 protocols using heparin salt solution
1
Tau Aggregation Kinetics Assay
2
Tau Aggregation Kinetics Assay
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Aggregation kinetics were measured in Synergy H1 microplate reader (BioTek) in black, flat-bottom, 96well plates (Nunc).Tau or tau 4R variants (10 μM) were pre-incubated in the presence or absence of indicated chaperones for 10 minutes at 37 °C. All proteins in the assay were buffer exchanged into the assay buffer (50 mM HEPES pH 7.4, 50 mM KCl, and 2 mM DTT). Thioflavin T (ThT; Sigma) at a final concentration of 10 μM was added and the aggregation was induced by the addition of a freshly prepared heparin salt solution (Sigma). Aggregation reactions were run at 37 °C with continues shaking (567 rpm) and monitored by ThT fluorescence (excitation = 440 nm, emission= 485 nm, bandwidth), using an area scan mode with a 3x3 matrix for each well. Black, flat-bottom, 96-well plates (Nunc) sealed with optical adhesive film (Applied Biosystems) were used. For data processing, baseline curves at same conditions but without heparin were subtracted from the data. Samples were run in triplicate and the experiments were repeated at least 4 times with similar results.
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