Applications of ZrO2 in the process for concentrating and purifying phosphorylated peptides
A technique for phosphorylated peptides and enrichment, applied in the field of enrichment and purification of phosphorylated peptides in the phosphorylated proteome
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
preparation example Construction
[0028] ZrO 2 The preparation of nanomaterials is obtained by co-precipitation method. The method is to use ZrOCl 2 The soluble zirconium salt and the precipitating agent ammonia are co-precipitated in the solution, and then calcined at high temperature to obtain ZrO 2 nanomaterials. This preparation of ZrO 2 The method of nanomaterials is simple, but the obtained nanomaterials have the problem of agglomeration. This can be obtained from figure 1 It can be seen in the scanning electron microscope image. Obtained ZrO 2 The size of nanoparticles is between 20-30nm.
[0029] ZrO 2 Synthesis of Nanomaterials
[0030] At room temperature, maintain pH = 10.0 under slow stirring to ZrOCl 2 .8H 2 O slowly add 25% NH 3 .H 2 O, get Zr(OH) 4 The precipitate was washed with double distilled water until the chloride ions were completely removed. Washed Zr(OH) 4 Drying at 110°C for 12 hours followed by calcination at 550°C for 4 hours yielded ZrO 2 Nanoparticles, scanning el...
Embodiment 1
[0039] 1. Preparation of sample solution: 1 mg of α-casein and β-casein were respectively dissolved in 1 mL of 50 mM ammonium bicarbonate solution (pH 8.2), according to the mass ratio with trypsin 1: 40 (w / w ) ratio to add trypsin for enzymolysis reaction, the reaction time is 16h, and the enzymolysis temperature is controlled at 37°C. The obtained proteolysis solution was stored in a -30°C refrigerator for future use.
[0040] 1mg of protein bovine serum albumin and ovalbumin were respectively dissolved in 1mL of reducing solution ((pH 8.2, 8M urea, 50mM ammonium bicarbonate solution), placed at room temperature for 4h, and then centrifuged into the centrifuge containing the inactivated protein Add DTT solution to the tube, and place it in a water bath at 37°C for 2 hours. Take the solution out of the water bath, add IAA solution to each tube solution, and let it stand in the dark at room temperature for 30 minutes. After the above treatment, the protein is completely denatu...
Embodiment 2
[0052] The difference from Example 1 is that
[0053] 2. ZrO 2 Nanomaterial-specific enrichment and purification of phosphorylated peptides
[0054] (1) β-casein, α-casein and Ovalbumin, which are hydrolyzed products of standard phosphorylated proteins, were respectively prepared in 10% HAC (pH=2-3) as the loading solution.
[0055] (5) Add 10 μL of 25% NH 3 .H 2 O Ultrasonic cleaning of ZrO 2 The nanoparticles were centrifuged at 30,000 rpm for 30 minutes, and the supernatant was collected.
[0056] Analysis results: Phosphorylated peptides from phosphorylated proteins α-casein, β-casein and ovalbumin digested by ZrO 2 Nanomaterials capture, but non-phosphorylated peptides are eluted, indicating that ZrO 2 Nanomaterials can specifically enrich and purify phosphorylated peptides. The phosphorylated peptides from the enzymatic hydrolysis products of phosphorylated protein β-casein and non-phosphorylated protein bovine serum albumin were treated with ZrO 2 Captured by na...
PUM
Property | Measurement | Unit |
---|---|---|
size | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com