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Osteoblast differentiation gene detection kit

A technology of osteoblast differentiation and gene detection, which is applied in the field of osteoporosis, can solve the problem of no research on the role of osteoblasts, and achieve the effect of high sensitivity

Inactive Publication Date: 2016-09-21
SHANGHAI INST FOR ENDOCRINE & METABOLIC DISEASES +1
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  • Abstract
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Problems solved by technology

Previous studies have shown that TFEB is closely related to the differentiation and function of osteoclasts, but its role in osteoblasts has not been studied

Method used

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  • Osteoblast differentiation gene detection kit
  • Osteoblast differentiation gene detection kit
  • Osteoblast differentiation gene detection kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1. Isolation and extraction of total RNA

[0035] 1) Discard the cell culture medium, rinse with PBS (1x) three times, add 1ml Trizol to each well of the six-well culture plate, and after lysis for about 5 minutes, blow the bottom of the plate with a 1ml pipette gun to suspend the adherent cells in Trizol , collect the Trizol cell suspension into a 1.5ml EP tube, store in a -80°C refrigerator or directly separate and extract;

[0036] 2) Add 200ul of chloroform to 1ml Trizol, close the cap of the EP tube tightly, invert the EP tube for 15 times and let it stand at room temperature for 15 minutes after the liquid becomes cloudy;

[0037] 3) Then put the EP tube after standing still in a low-temperature centrifuge, centrifuge at 12000 rpm for 15 minutes at 4°C. Carefully take out the EP tube, it can be seen that the solution in the EP tube is stratified, the lower layer is a red phenol-chloroform solution containing DNA; the thin middle white layer contains protein; the ...

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Abstract

The invention relates to an osteoblast differentiation gene detection kit. The kit contains primer pairs of Atg5, Atg7, BECN1, LC3, ALP, OC, CollengeI and OPG. The osteoblast differentiation gene detection kit has high detection sensitivity; as proved by detection of the kit, the change of an autophagy level in an osteogenic differentiation process is verified, and convenience is brought to personal treatment, prognosis and preventive intervention of clinical patients.

Description

technical field [0001] The invention belongs to the field of osteoporosis, in particular to a kit for detecting osteoblast differentiation genes. Background technique [0002] The growth and development of bone are closely related to the function of osteoblasts. Osteoblasts start the process of bone formation by secreting a large amount of collagen and bone matrix, as well as some important cytokines and enzymes, and couple with the regulation of osteoclasts through a series of factors to control the formation and maturation of osteoclasts and activation, so that bone formation and bone resorption are in a dynamic balance. The main pathological features of osteoporosis are the loss of bone mass and the degeneration of bone microstructure caused by the uncoupling of bone formation and bone resorption. Fractures and fracture-related complications caused by osteoporosis impose a huge economic burden on society and families. At present, there are many kinds of drugs for the t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q2600/118C12Q2600/158
Inventor 宁光赵红燕赵点点孙立昊陶蓓刘建民
Owner SHANGHAI INST FOR ENDOCRINE & METABOLIC DISEASES
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