BTA-2

BTA-2, a benzothiazole dye, is structurally similar to thioflavin T (ThT), which exhibits an enhanced fluorescence signal when bound to amyloid fibrils. BTA-2 has distinct absorption and emission characteristics in solution and when bound to amyloid fibrils, which makes it can used for identifying amyloid fibrils using spectroscopy[1].

Research Areas >> Others

Signaling Pathways >> Others >> Others

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs). To research how CH3CN affects binding of amyloid fibrils (varied the amount of CH3CN but maintained the same concentration of BTA-2, fibrils, and water) and how the concentration of BTA-2 affects binding (varied the amount of BTA-2 but had a constant amount of CH3CN, fibrils, and water) [1] . BTA-2 in solution and in the presence of amyloid fibrils: 1. Insulin obtains from a bovine pancreas, and is stored at −20°C in a desiccator. 2. To synthesize BTA-2 and further purified. 3. To prepare fibrils: Insulin is dissolved in pH 2 water (5 mg/mL) and filters through a 0.2 μm filter, heats at 60℃ for 24 h. Centrifuge at 3000 rpm for 2.5 min to remove any globular artifacts. The supernatant containing the fibrils is saved for later use. Solutions are stored in a refrigerator (10℃) until needed. 4. BTA-2/fibrils solutions (are prepared in two ways): (First way) Diluting (100 μL) of the stock fibril solution with 880 μL of pH 2 water, adding (20 μL) of 0.8 mg/mL (3.15 mM) BTA-2 in CH3CN. (Second way) Diluting (100 μL) of the stock fibril solution with a 1:1 mixture of nanopure water: CH3CN, adding (20 μL) of 0.8 mg/mL (3.15 mM) BTA-2. 5. Two similar solutions are made that contained only the BTA-2 without the fibrils. 6. The standard amyloid fluorescent marker is thioflavin T (ThT): 20 μL of 0.8 mg/mL (2.51 mM) ThT in CH3CN and 100 μL of stock fibril solutions is diluted with 880 μL of pH 2 water.7. The emission spectra of BTA-2 in pH 2 water excited at 430 nm and excited at 360 nm, respectively.

[1]. Catherine C Kitts, et al. A spectroscopic study of 2-[4'-(dimethylamino)phenyl]-benzothiazole binding to insulin amyloid fibrils. J Fluoresc. 2010 Jul;20(4):881-9.