Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Preparation of efficient and stable trace kit for detecting triglyceride in fat cells

A detection kit and adipocyte detection technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of easily damaged cell layers, high absorbance, oil red residue, etc., and reduce the experimental time and operation steps Simple, high-throughput effects

Active Publication Date: 2016-02-03
SICHUAN AGRI UNIV
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the rapid development of cell experiment technology, the requirements for experimental accuracy are getting higher and higher, and the classic oil red O staining method has also exposed some of its own defects, such as: cells need to be fixed and washed repeatedly during staining, which takes a long time and Easy to damage the cell layer; oil red O working solution needs to be filtered before use, otherwise there will be oil red residue after staining, which will affect the observation; it is not easy to wash off the excess dye solution after staining, resulting in high absorbance detected after isopropanol extraction ;Saturated Oil Red O liquid is stored for too long, and the color will become dark black, which will affect the use; During long-term operation, the volatilized isopropanol will affect the health of the experimenter

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Preparation of efficient and stable trace kit for detecting triglyceride in fat cells
  • Preparation of efficient and stable trace kit for detecting triglyceride in fat cells
  • Preparation of efficient and stable trace kit for detecting triglyceride in fat cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0027] Preadipocytes were isolated and purified from the subcutaneous adipose tissue of the neck and back of Tibetan pigs 2 days after birth, and differentiated by cocktail method (Insulin, Dex, IBMX) + rosiglitazone, and the cells on day 12 after differentiation were used for the experiment. No. 5 formula of the present invention is used in the experiment, and the detailed steps are as follows:

[0028] Step 1: Induce the differentiation of Tibetan pig preadipocytes in a 96-well plate for 12 days, take out the plate from the incubator, carefully suck off the culture medium, and pay attention not to touch the underlying cells with the tip of the pipette;

[0029] Step 2: Wash twice with PBS (200ul / well), as gently as possible, after washing, add 200ulPBS to each well;

[0030] Step 3: Add 5ul Nile Red working solution and incubate at room temperature (20-25°C) in the dark for 5 minutes;

[0031] Step 4: Place the orifice plate in a microplate reader, and set a wavelength of 4...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a kit for rapid detection of content of triglyceride in fat cells. The kit mainly consists of a Nile red working solution which takes acetone and DMSO as solvents, and PBS. The invention also provides an intracellular triglyceride fluorescent staining method. Compared with a conventional oil red O staining method, an operating method of the kit is simpler, short in time consumption, high in efficiency, high in flux, low in content of impurities and is easier for users to observe, and the operating method is capable of quantifying triglyceride in fat cells within a short time. Therefore, the kit disclosed by the invention is more suitable for current fat cell differentiation researches.

Description

technical field [0001] The invention relates to a trace, efficient and stable triglyceride detection kit in adipocytes, which belongs to the technical field of biological reagents. Background technique [0002] In vitro, preadipocytes are induced (insulin, dexamethasone, IBMX, etc.) to differentiate into mature adipocytes. During the differentiation process, triglycerides are produced in the cytoplasm of the cells and gradually accumulate to form round lipid droplets (lipiddroplets). The number and degree of accumulation of lipid droplets can be used as phenotypic values ​​to assess the degree of differentiation of preadipocytes. Currently, lipid droplets in mature adipocytes are quantified and qualified by staining with lipophilic dyes and extracting to detect absorbance. [0003] Oil Red O can stain lipid droplets in mature adipocytes red, and at the same time, the amount of triglycerides can be quantified by detecting the absorbance after extraction with isopropanol. Th...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12Q1/61
Inventor 张进威王讯罗毅王鹏俊李明洲
Owner SICHUAN AGRI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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