Method for intracellular delivery of protein
A protein and protein solution technology, applied in the field of bioengineering, can solve problems such as easy to be removed, protein cannot function, CPPs short half-life, etc., to achieve the effect of avoiding chemical bond modification, excellent biocompatibility, and simple operation steps
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0023] (1) Take boron halide clusters (Na 2 B 12 Br 12 ) and labeled cytochrome C, the solvent is 1×PBS, the total volume is 100 μL, mixed thoroughly for 10 min, and sterilized by filtration through a 0.22 μm microporous membrane;
[0024] (2) Add 300 μL DMEM to the 24-well plate where NIH-3T3 cells were cultured, and add 100 μL of the mixture in step (1), incubate for 6 hours, boron halide clusters (Na 2 B 12 Br 12 ) with a final concentration of 150 μM, and labeled cytochrome C concentrations of 2.5 μM, 5.0 μM and 10.0 μM;
[0025] (3) Aspirate the mixture in the 24-well plate in step (2), wash with 1×PBS three times, stain and fix the cells;
[0026] (4) Carry out confocal imaging of the cells using a confocal microscope to observe the distribution of labeled cytochrome C in the cells. The distribution of proteins in the cells can be seen in the fluorescent channel of 500-530nm labeled cytochrome C, from figure 2 Boron halide clusters (Na 2 B 12 Br 12 ) can effici...
Embodiment 2
[0028] (1) boron halide clusters (Cs) with a mass ratio of about (1~5):1 2 B 10 Br 10 ) and labeled cytochrome C, the solvent is 1×PBS, the total volume is 100 μL, mixed thoroughly for 10 min, and sterilized by filtration through a 0.22 μm microporous membrane;
[0029] (2) Add 300 μL DMEM to the 24-well plate where NIH-3T3 cells were cultured, and add 100 μL of the mixture in step (1), incubate for 6 hours, boron halide clusters (Cs 2 B 10 Br 10 ) with a final concentration of 150 μM, and labeled cytochrome C concentrations of 2.5 μM, 5.0 μM and 10.0 μM;
[0030] (3) Aspirate the mixture in the 24-well plate in step (2), wash with 1×PBS three times, stain and fix the cells;
[0031] (4) Carry out confocal imaging of the cells using a confocal microscope to observe the distribution of labeled cytochrome C in the cells. The distribution of proteins in the cells can be seen in the fluorescent channel of 500-530nm labeled cytochrome C, from image 3 boron halide clusters (C...
Embodiment 3
[0033] (1) Take boron halide clusters (Na 2 B 12 Br 12 ) and labeled cytochrome C, the solvent is 1×PBS, the total volume is 100 μL, mixed thoroughly for 20 minutes, and sterilized by filtration through a 0.22 μm microporous membrane;
[0034] (2) Add 300 μL DMEM to the 24-well plate in which HeLa cells were cultured, and add 100 μL of the mixture in step (1), incubate for 6 hours, boron halide clusters (Na 2 B 12 Br 12 ) final concentration is 200 μM, and the concentration of labeled cytochrome C is 1.0 μM;
[0035] (3) Aspirate the mixture in the 24-well plate in step (2), wash with 1×PBS three times, stain and fix the cells;
[0036] (4) Use a confocal microscope to perform confocal imaging of the cells to observe the distribution of the labeled cytochrome C in the cells. The fluorescent channel of the labeled cytochrome C at 630-670nm can be used to see the distribution of proteins in the cells. Flow cytometry The instrument detects the efficiency of labeled cytochro...
PUM
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