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细胞培养之细胞复苏小妙招
2023-01-11
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原代细胞分离提取方法及注意事项
2022-11-30
原代细胞是直接取自活组织的细胞,并用于体外生长。这些细胞经历了非常少的群体倍增,因此与连续(肿瘤或人工永生化)细胞系相比,它们更能代表它们所衍生组织的主要功能成分,使得原代细胞成为更具代表性的体内模型。
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核酸提取
2022-11-30
核酸提取是指将核酸(DNA和RNA)从细胞中提取出来的过程,总的来说包括细胞裂解、核酸纯化及后续核酸浓度、纯度测定的步骤。根据原始样品的种类和核酸产物后续的各种应用目的,细胞裂解和核酸纯化步骤有不同的处理方法和要求。
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核酸检测
2022-11-30
核酸浓度与纯度检测最简单常用的仪器是微量紫外分光光度计,代表为NanoDrop。NanoDrop分光光度计利用分子的紫外吸收特性来测定样品浓度。核酸吸收峰为260nm,蛋白样品吸收峰为280nm。此外,很多提取过程中残留的污染物通常在280nm和230nm处有吸收峰。[ NanoDrop Spectrophotometer Resources | Thermo Fisher Scientific - CN]样品浓度与吸光度成正比关系(Beer-Lambert Law),因此通过测定吸收峰就可以得出核酸样品的浓度。
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单克隆抗体
2022-11-23
抗体是脊椎动物为应对外来入侵蛋白(抗原),保护机体不被伤害而分泌的、在体内进行循环的蛋白质分子,是机体体液免疫的主要参与者。抗体又被称为免疫球蛋白(immunoglobulin,Ig),通过电泳技术对血清蛋白进行精确的分析,发现大部分的抗体蛋白都集中在伽马球蛋白(gamma globulin)所在的条带,因此抗体也被称为免疫球蛋白。[血清蛋白通过物理的溶解度定义,可以被简单的区分为白蛋白(albumin)和球蛋白(globulin)]
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培养基配置与储存
2022-11-23
根据配置体积,选择合适的灭菌后容器,在容器中倒入约90%体积的超纯水,将培养基干粉全部倒入该容器中,用少量超纯水将袋内残留粉末洗出。
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细胞侵袭和迁移
2022-11-02
实验准备: 细胞,24孔板,细胞小室(8μm,24孔板专用),ECM Gel,10%FBS+培养基,无血清培养基,10 μ、200 μL、1000 μL移液器及配套枪头,1.5 mL EP管,冰盒
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细胞冻存与复苏
2022-11-02
一、如何处理收到的细胞 1.新买的或惠赠的细胞如何处理? 不管买的细胞、惠赠的细胞,刚拿到手应赶紧做这几件事:检测瓶子是否破裂;培养基是否漏出,是否浑浊;是否有支原体污染;迅速扩增冻存。
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