Fluorescent probe for detecting Abeta oligomers as well as preparation method and application thereof
A fluorescent probe and oligomer technology, applied in fluorescence/phosphorescence, chemical instruments and methods, luminescent materials, etc., can solve problems such as unclear strategy and few fluorescent probes, and achieve simple post-processing process and high sensitivity. High, easy-to-operate effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0069] Embodiment 1 synthetic organic small molecule fluorescent probe (I)
[0070] The synthesis of a kind of organic small molecule fluorescent probe (I) of the present embodiment, the synthesis route diagram is as follows figure 1 As shown, the specific synthesis steps are as follows:
[0071] (1) Synthesis of Intermediate 1
[0072] ① Mix the reactant p-hydroxybenzaldehyde (0.02mol, 2.44g), N-acetylglycine (0.02mol, 2.34g), anhydrous sodium acetate (0.018mmol, 1.47g) and acetic anhydride 20mL, then heat and stir for 110 Reflux at ℃ for 4h, after the reaction is completed, slowly add heated (50℃) ethanol, the reaction liquid is cooled to room temperature, cooled for 4h, crystals are precipitated, filtered, washed with ice ethanol, and then washed with hot water to obtain a yellow solid, dried to obtain 3.64 g.
[0073] ② Mix yellow solid (0.01mol, 2.03g), potassium carbonate (0.015mol, 2.07g), 33% (v / v) methylamine aqueous solution 4mL and 4mL ethanol in a 75mL volumetri...
Embodiment 2
[0082] The ultraviolet-fluorescence test of embodiment 2 organic small molecule fluorescent probe (I)
[0083] The probe (I) prepared in Example 1 was dissolved in dimethyl sulfoxide (DMSO) to prepare a 10 mM mother solution; then 3 μL of the 10 mM mother solution was dissolved in 3 mL of DMSO to obtain a 10 μM probe solution. Test the UV absorption spectrum under the UV spectrophotometer and record the maximum absorption wavelength. Test the fluorescence spectrum under a spectrofluorometer and record the maximum emission wavelength.
[0084] The result is as Figure 5 Shown in A: 10 μM probe (I) has a maximum absorption wavelength of 546 nm, a maximum emission wavelength of 650 nm, and a Stokes shift of 104 nm in DMSO solution.
Embodiment 3
[0085] Embodiment 3 The evaluation of the binding ability of organic small molecule fluorescent probe (I) and Aβ protein
[0086] Fluorescence titration spectrometry: add an appropriate amount of 100 μM Aβ protein (Aβ monomer, Aβ oligomer and Aβ aggregate, purchased from Shanghai Qiangyao Biology) solution in PBS buffer (pH=7.4, 10mM), the total volume is 600 μL, keeping the concentration of probe (I) (prepared in Example 1) at 1 μM and the protein concentration at 10 μM, and measuring its fluorescence spectrum under a fluorescence spectrophotometer for data processing. The parameters of the fluorescence spectrophotometer are: working voltage 700V, outer slit=10nm, electromagnetic slit=10nm.
[0087] The result is as Figure 5 Shown in B: Compared with Aβ monomers and Aβ aggregates, after the probe (I) binds to Aβ oligomers, the fluorescence intensity increases significantly.
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More