哺乳动物细胞蛋白抽提试剂 SMC Mammalian Cell Protein Ext
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·哺乳动物细胞蛋白抽提试剂说明
The SMC Mammalian Cell Protein Extraction Reagent from SinoBio, containing nonionic, nondenaturing detergent, is a reliable cell lysis buffer used to lyse mammalian cells from grown in both adherent and suspension cultures. It is compatible with protein assays, immunoassays and protein purification. SMC Protein Extraction Reagent does not contain protease or phosphatase inhibitors. If needed, protease inhibitors and phosphatase inhibitors can be added to the Protein Extraction Reagent just before use to prevent proteolysis and maintain phosphorylation of proteins.
· 哺乳动物蛋白抽提试剂使用方法
Protocol for Lysis of Mammalian Cells Grown in Suspension Culture
1. Pellet the cells by centrifugation at 2500rpm for 5 minutes at 4°C.
2. Aspirate the supernatant.
3. Wash the cells twice with cold PBS.
4. Add 1 ml of SMC Mammalian Cell Protein Extraction Reagent per 5x106-1x107 cells to the cell pellet.
5. Resuspend cells by pipetting up and down. Vortex cells for 10 seconds at maximum speed.
6. Shake the mixture on ice for 15 minutes.
7. Centrifuge the lysate at 14,000 rpm for 10 minutes at 4°C.
8. Transfer the supernatant to a new tube.
Protocol for Lysis of Mammalian Cells Grown in Adherent Culture
1. Aspirate the media.
2. Wash the cells twice with cold PBS. Remove the PBS as much as possible.
3. Add an appropriate volume of SMC Mammalian Cell Protein Extraction Reagent.
Culture vessel Volume of Protein Extraction Reagent
100 mm dish 500-1,000 μl
60 mm dish 250-500 μl
35 mm dish 200-400 μl
6-well plate 200-400 μl per well
24-well plate 100-200 μl per well
96-well plate 50-100 μl per well
T-25 flask 250-500 μl
T-75 flask 500-1,000 μl
4. Incubate for 20 minutes at 4°C with shaking.
5. Use a cell scraper to gather the lysate and transfer everything to a 1.5 ml tube.
6. Centrifuge tube at 14,000 rpm for 10 minutes at 4°C.
7. Transfer the supernatant a new tube.
·其他说明
详细内容请参考说明书
