原代培养

A general protocol for staining cell for cytometry analysis General Annexin V Staining Procedure Solutions 1.10X Binding Buffer (Cat. No. 66121A):0.1 M HEPES pH 7.4; 1.4 M NaCl; 25 mM CaCl 2 . Di ...

When an irreplaceable culture becomes contaminated researchers may attempt to eliminate or control the contamination. First determine if the contamination is bacteria fungus mycoplasma or yeast. Isola ...

Authentication Techniques Whatever the scope of work to be carried out it is important to know that the work is being conducted using the correct reagents. This is no less important for cell cultures ...

We use two different kinds of media. Most cells are grown in DMEM. A few lymphoid cell lines are grown in RPMI. Cells grown in DMEM must be grown in a 10% CO2 atmosphere. As a result most of our incub ...

1. embryos are dissected from timed-pregnant mice from 11.5 d.p.c. to 13.5 d.p.c. 2. metanephroi and associated ureteric buds are microdissected and placed in holding medium (L15 medium supplemented ...

embryos are dissected from timed-pregnant mice from 10.5-11.5 d.p.c. limb buds are microdissected and placed in holding medium (L15 medium supplemented with 1 x MITO+ serum extender) (all me ...

Isolation of Lung Bud Endoderm What you need: E11-12 mouse embryos DMEM with 5% fetal bovine serum petri dishes for dissections and washes Tyrode-Ringer's solution pH 7.6-7.7 (recipe below) pancr ...

Phosphate-buffered saline (PBS) (0.01 M Na2HPO4 0.15 M NaCl pH 7.4):sodium phosphate dibasic (Na2HPO4) 0.87 gsodium phosphate m ...

Healthy Sp2/0 cells should be rapidly growing by this time. Sp 2/0 cells should be started about two weeks before the cell fusion. Every two days they should be centrifuged at a 64.4 xg on the IEC cli ...

Protocol is for 150 ml of cell culture. 1. Spin down cells and wash 1X in 10 ml ice cold PBS. 2. Resuspend pellet in 20 ml lysis buffer: 50 mM Tris pH 8.0 1 % NP-40 ...

293细胞是腺病毒载体的包装细胞。腺病毒是继逆转录病毒后用于基因治疗研究的热门载体。直接将腺病毒载体导入人体内表达目的基因，治疗恶性肿瘤，心血管疾病或一些遗传疾病已取得可喜进展；利用腺病毒载体在包装细胞 293中表达分泌性蛋白质，如蛋白酪氨酸激酶 1C等亦成为生产重组蛋白的一条途径。因此完善 293细胞的大规模培养技术具有越来越重要的市场意义。哺乳细胞的大规模培养方式有三种：贴壁培养，微载体培养， ...

Day -1 Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells. Day 1 Trypsinized the cells as for normal passaging until the colonies li ...

Day -1 Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells. Day 1 Trypsinized the cells as for normal passaging until the colonies li ...

Day -1 Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells. Day 1 Trypsinized the cells as for normal passaging until the colonies li ...

1. Treat 10 cm tissue culture plates with 0.1% gelatin for at least two hours before use. 2. Sacrifice the pregnant female mouse (day 13 or 14 p.c.) by cervical dislocation. Dissect out the uterine h ...

Three weeks prior to embryonic fibroblast isolation a PGK-neo male mouse is mated to a heterozygote female. 13.5 days to 14.5 days after the plug is observed sacrifice the pregnant female either by ce ...

细胞培养常见问题及解答

The technique described here is a slight modification (March 1997) of methods presented in: Nagy A. J. Rossant. 1993. Production of completely ES cell-derived fetuses. In : Gene Targeting: A Practical ...

Day 0 One frozen vial of Murine Embryonic Fibroblasts (MEFs) is thawed quickly in a 37oC water bath. When the last bit of ice is melted spray the vial with 70% ethanol and transfer the contents of the ...

1.Ice on tray.2. Dissection tools: 1 pair of large scissors; 2 pairs of fine scissors; 1 pair of coarse forceps; 2 pairs of fine forceps; soak in 95% EtOH.&nb ...