细胞技术

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 ...

血清的热灭活是很多培养者感兴趣的话题，价格不菲的血清中含有诸如生长因子，维生素，氨基酸等珍贵物质，而将它们置于50℃以上的温度长达30分钟是完全没有必要的。尽管如此，在大多数实验室之中血清的热灭活还是作为常规来执行，多数实验者并没有考虑热处理对血清中的生长因子，氨基酸等成分带来的负面影响，在我们的技术热线中最常被提到的就是否该对血清进行热灭活，下面我们就对胎牛血清的热灭活进行一些探讨和解释。

培养基DMED的配置。

细胞原代和传代培养的步骤及注意事项。

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 ...

细胞差异和分化，细胞生长和增殖。

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 ...

以下是根据NIH老年病研究所提供的英文的R1胚胎干细胞培养protocol整理而成。根据经验作部分修改。 1、一般培养：保持胚胎干细胞处于未分化状态 培养基细胞复苏冻存细胞明胶包被&n ...

Passaging cells Pour out media from flasks. Wash with Hanks. 5 ml per flask. Tilt around then dump. Add 4 ml of Trypsin / EDTA to each flask. Tip then bang. Add 20% FBS NCTC media and tilt. 4 ml p ...

Terminal deoxynucleotidyl Transferase-mediated dUTP nick end labeling (TUNEL) is an in situ method for detecting the 3'-OH ends of DNA exposed during the internucleosomal cleavage that occurs during a ...

Protocol for Paraffin Sections: Dewax paraffin sections: Incubate slides 55°C 30 min. Xylenes 2 times 2 min. each 100% EtOH 2 times 2 min. each&nb ...

Protocol I: Triton X-100 Lysis Buffer In 96 flat-wells plate incubate 4x10 6 target cells (40 wells of 105 per well) with desired concentration of effectors (105 target cells per well). After incubat ...

Materials from a workshop held March 1997 in Modena Italy. All materials reproduced with permission. Introduction A. Cossarizza Workshop Sponsors Methods in analysis of apoptosis and cell necrosis. Z. ...

BrdU Incorporation Protocol Day 1 Label cells with media containing100 μM BrdU (6 μL/mL of 5 mg/mL stock): Diploid Fibroblasts 4-6 hours ES cells 30 minutes 3T3 cells 2.5 hours Trypsinize ce ...

immunofluorescence labeling; that is the antibodies have the fluorescent dye attached. Direct labeling is simpler and quicker than indirect labeling. We strongly recommend direct labeling if you're pl ...

FIXATION and DNA Staining for Cell Cycle Analysis BackgroundThis method of DNA staining utilizes ethanol to fix the cells and permeabilize the membrane which allows the dye (Propidium Iodide) to enter ...