Quick Overview for Osteoclast (大鼠) Culture Kit (ABIN956046)
抗原
Osteoclast
适用
大鼠
应用范围
Cell Culture (CC)
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组件
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2x Rat Osteoclast Precursor Cells, frozen. Vial containing 2 x 10^6 cells
1x Washing Medium, alpha-MEM. 50 mL
1x Culture Medium, alpha-MEM with M-CSF (10 ng/mL) and RANK Ligand (10 ng/mL). 25 mL -
试剂未包括
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Pipettes
96-well, flat bottom culture plate
Tubes
Refrigerated centrifuge
Water bath.
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结果分析
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- Thaw a vial of primary precursor osteoclasts in a 37°C water bath.
2. After thawing, transfer the cells to a 15 mL centrifuge tube, add 10 mL of Wash Medium and mix briefly. Centrifuge 1,000 rpm for 5 minutes at 4°C.
3. Remove supernatant and add 10 mL of Wash Medium and mix briefly. Centrifuge 1,000 rpm for 5 minutes at 4°C.
4. Remove supernatant and resuspend the cells in 2.5
5 mL of Culture Medium. To study factors that effect osteoclasts formation, add the factors to the Culture Medium.
5. Transfer 100 µL of cell suspension into each well of a 96-well plate. If the cells are resuspended in 5 mL of Culture Medium, there will be enough cell suspension for about 50 µLs. To quickly observe osteoclasts formation, culture the cells at a higher density.
6. Feed the cells with 100 µL of Culture Medium every 3
4 days. Cells will begin to fuse and form osteoclasts around day 5 (fig 1).
7. Count the osteoclasts by staining with tartrate-resistant acid phosphatase (TRAP Staining Kit,.
- Thaw a vial of primary precursor osteoclasts in a 37°C water bath.
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限制
- 仅限研究用
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注意事项
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1. Read the instructions carefully before beginning the culture.
2. This kit is for research use only, not for human or diagnostic use.
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- Osteoclast
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背景
- In aging societies, osteoporosis and other age-related bone metabolism disorders are a rapidly increasing problem. The amount of bone in an organism is controlled by a balance of osteoblasts (bone-forming cell) and osteoclasts (bone- destroying cell) activities. A method to induce osteoclasts formation from bone marrow cells using M-CSF (macrophage- colony stimulating factor) and RANKL (receptor activator of NF-B ligand) has been established in recent years. This kit includes cryopreserved primary precursor osteoclasts from rat bone marrow and Culture Medium containing M-CSF and RANKL.
抗原
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