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1.当细胞铺满整个平皿(100mm)后,吸弃旧培养液,10mLPBS洗一遍
2.0.25%胰酶(预热20分钟左右)2ml消化,轻轻摇晃,Osteocalcin,affinity purified (human) From acid demineralized bone extracts直至肉眼见单层细胞呈块状脱落(或在显微镜下观察细胞之间分离),立刻加5ml(DMEM+10%FBS)终止消化,轻柔吹打8-10次成单细胞。
3.离心1000rpm*5min,弃上清,加DMEM+10%FBS轻柔吹打。
4.将细胞悬液1:4传到100mm细胞培养皿中,每个皿加10ml(DMEM+10%FBS)培养液,正常情况下2-3天就可以长满,期间不用换液。
Osteocalcin,affinity purified (human) From acid demineralized bone extracts的传代方法:首先在倒置显微镜下观察生长融合达80%,不需要长满,就准备传代。倒掉旧的培养基,再用已经高压灭菌过的PBS洗1-2次,我用的是25平方厘米的培养瓶,加入1毫升已经配制好的0.25%胰蛋白酶-0.02%EDTA(0.25%胰蛋白酶是用D-Hanks配制后先用滤纸过滤,再用一次性过滤器过滤除菌,Osteocalcin,affinity purified (human) From acid demineralized bone extracts再分装成1ml的小包装,刚好每次用完一个,避免每次反复冻存,会使胰酶的活性下降),加入的量以刚好盖满瓶底为宜。放到倒置显微镜下观察变化,一旦发现细胞开始变园收缩,大约1-3分钟时间,必要时可以吹打帮助细胞分散,就立即加入培养基中和胰酶,如有EDTA最好要离心(800rpm,5min),弃上清夜,Osteocalcin,affinity purified (human) From acid demineralized bone extracts再加入完全培养基吹匀,再分瓶,一般一瓶可以分2-3瓶。放入5%的CO2培养箱,37度培养24小时后观察。
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xyA387Hu 补体3a(C3a)ELISA试剂盒 检测范围
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文献和实验Protein A Mimetic (PAM) Affinity Chromatography: Immunoglobulins Purification
of 4�C, and the room temperature-dependent elution of adsorbed immunoglo bulins in the presence of ethylenediaminotetraacetic acid (EDTA). This ligand shows low binding affinity for IgG, but binds to bovine and human IgM with reduced affinity than murine
interactions were evaluated at the C/EBP site in the rat Osteocalcin promoter (A) . Oligonucleotides (30 bp) representing wild-type (WT) and mutant (MT) sequences for C/EBP site were gel purified. WT probe was incubated with 6 μg of HeLa nuclear extracts
Methods for the Measurement of a Bacterial Enzyme Activity in Cell Lysates and Extracts
were studied in situ in cell-free extracts to obtain information on enzyme function in an environment that was closer to the bacterium's physiological conditions. There are numerous and unique advantages in studying enzyme activities in purified preparations
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