Magnetic carbon sphere of surface finished C8 alkyl chain, preparation and application thereof
A technology of surface modification and alkyl chains, applied in the field of magnetic carbon spheres and its preparation, can solve problems such as cumbersome operations, and achieve good practical value and application prospects
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Embodiment 1
[0026] Embodiment 1, the synthesis of the magnetic carbon sphere of surface modification C8 alkyl chain
[0027] The synthesis of magnetic carbon spheres with surface-modified C8 alkyl chains is divided into three steps.
[0028] First, the aminoferroferric oxide magnetic nanoparticles were synthesized by hydrothermal method: 1.0 g FeCl 3 ·6H 2 O was dissolved in 30 mL of ethylene glycol, and magnetically stirred for 0.5 h to obtain a yellow transparent solution. Then 4.0 g of anhydrous NaAc was added, and after magnetic stirring for 0.5 h, a brown-yellow transparent solution was obtained. The resulting solution was transferred into a 200 mL Teflon-lined stainless steel reaction kettle. Put it in an oven at 200°C for 12 hours. After cooling to room temperature, the product was repeatedly washed with deionized water five times to remove water-soluble impurities such as sodium acetate and ethylene glycol, and finally the product was vacuum-dried at 50°C for future use.
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Embodiment 2
[0031] Example 2, Magnetic carbon spheres with surface-modified C8 alkyl chains used for peptide enrichment
[0032] (1) Enzymatic hydrolysis of protein solution:
[0033] Take 1.0 mg protein sample, including bovine serum albumin (BSA), cytochrome C (Cytochrome C), horse myocardium (Myoglobin), egg albumin (Ovalbumin), bovine β-casein (β-casein) and milk The extracted casein (Casein) was dissolved in 1.0mL water respectively, and after denatured by heating, ammonium bicarbonate solution was added to the solution to adjust the pH of the system to about 8, and 25 μg trypsin. At a temperature of 37°C, the enzymatic hydrolysis was terminated after 12 hours, and the enzymatic hydrolysis solution was frozen in a -80°C refrigerator until use.
[0034] (2) Separation and enrichment of standard peptides:
[0035] Make 10 μL a concentration of 5 mg mL -1 The magnetic material dispersion solution was added to 400 μL with a concentration of 5 fmol μL -1 In standard peptide aqueous s...
Embodiment 3
[0044] Embodiment 3 surface modification Fe of C8 alkyl chain 3 o 4 @SiO 2 Magnetic material used for enrichment of enzymatic hydrolyzate on gel
[0045] First, the protein in the human lens was extracted, and 150 mg of the protein was separated by two-dimensional gel electrophoresis, and gel spots with a lower concentration were selected for enzymatic hydrolysis on the gel to obtain about 400 μL of enzymatic hydrolysis solution, which was stored at -20°C for later use. Make 10 μL a concentration of 5 mg mL -1 The magnetic material dispersion solution was added to 200 μL of the above enzymatic hydrolysis solution, and shaken at room temperature for 10 minutes. After removing the supernatant under the action of an external magnetic field, wash it repeatedly with pure water three times; then add 2 μL of 50% (v / v) acetonitrile aqueous solution, and spot 0.5 μL of the mixture containing the enriched peptides and magnetic microspheres onto the target plate After drying, MALDI-T...
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