Method for synthesizing poly-dopamine-modified MALDI target plate and application of poly-dopamine-modified MALDI target plate
A polydopamine and synthesis method technology, which is applied to measurement devices, material analysis by electromagnetic means, instruments, etc., can solve problems such as contamination and damage of mass spectrometry ion sources, avoid cumbersome elution, and the synthesis method is simple and good. The effect of biocompatibility and hydrophilicity
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Embodiment 1
[0029] Synthesis of polydopamine-modified MALDI target plate
[0030] (1) Disperse 400mg of dopamine hydrochloride in 200mL of Tris buffer (10mM, pH=8.5).
[0031] (2) Wash the MALDI target plate several times with distilled water and ethanol, and dry at room temperature.
[0032] (3) Submerge the cleaned MALDI target plate in the dopamine solution in (1) and react at room temperature for 24 hours.
[0033] (4) The polydopamine-modified target plate obtained in (3) was washed three times with distilled water, and immersed in Ti(SO 4 ) 2 (100 mM) aqueous solution for 2 hours at room temperature.
[0034] (5) Rinse the target plate obtained in (4) three times with distilled water and dry at room temperature.
[0035] figure 1 It is a scanning electron micrograph of the surface of the MALDI target plate modified by polydopamine. Dopamine presents a small spherical shape at the beginning of the polymerization reaction, and tends to form a polydopamine film at the later stag...
Embodiment 2
[0036] Example 2: Polydopamine-modified MALDI target plate selectively enriches phosphorylated peptides and performs high-throughput MALDI-TOF MS analysis
[0037] (1) Sample preparation: Dissolve BSA or β-casein protein in NH 4 HCO 3 buffer (25mM, pH 8.3) and digested with trypsin (2%, w / w) at 37°C for 16 hours. The product after enzymatic hydrolysis should be stored below 0°C. After the serum samples were centrifuged, the supernatant was stored below 0°C.
[0038] (2) Spotting the target: wash the modified target plate with 50% acetonitrile and 0.1% TFA aqueous solution (v / v) three times, respectively add the β-casein enzymatic hydrolysis solution, the mixture of β-casein and BSA enzymatic hydrolysis solution and The treated serum was diluted to different concentrations with 50% acetonitrile and 0.1% TFA aqueous solution (v / v); pipette 1 μL of the diluted enzymolysis solution onto the modified MALDI target plate, and place it in a wet box at room temperature for enrichme...
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