细胞功能测定

用荧光激活的细胞分类仪检测调亡细胞1）原位缺口翻译检测裂解的DNA片段1．取106经促调亡物质处理的细胞，用1％ BSA-PBS洗涤细胞后加入0.1ml FITC标记的抗CD4或抗CD8单克隆抗体，4℃ 20分钟。用1％ BSA-PBS洗涤细胞。置冰上。2．加入0.1ml 1％多聚甲醛，置冰上5分钟，加入60％（v/v）甲醇溶液，轻轻悬浮细胞，4℃固定细胞15分钟。4℃离心500×g 10分钟，去 ...

用淋巴细胞分离液分离单个核细胞1）分离外周血中的单个核细胞（PBMC）1．抽取正常人静脉血加到肝素抗凝管中，加等量含5～10IU/ml肝素的无血清缓冲液悬浮细胞。将细胞悬液小心加在与血液等量的淋巴细胞分离液上，室温中，水平离心500×g 20分钟。此时离心管中形成5层：最上面是血浆，血浆层和淋巴细胞分离液之间是PBMC，淋巴细胞分离液层和最下面的红细胞层之间是粒细胞层，又成为棕�层。2．吸去最上层 ...

METHOD 1:Formaldehyde preservative � 2% formaldehyde solution in protein-free phosphate-buffered saline (PBS).Prepare as follows:Add 2 g paraformaldehyde powder (e.g. Sigma St. Louis MO) to 100 ml of ...

准备工作： DF12加入0.1%BSA，100ml取10ml冰浴，40ml放入孵箱；其余的放入4度； DNaseI10undefined;PI：50ug/ml;Hoechst33342:1000ug/ml（allfromSigma） 计数板，冰盒；各种离心管，无菌流式管（BD），400滤网（BD） 染色方案： ①细胞染色： 细胞消化计数，用DF12重悬，调整到1×106/ml浓度 ...

上皮细胞培养 1）表皮细胞培养 1．取材：取外科植皮或手术残余皮肤小块，以角化层薄者为佳，早产流产儿皮肤更好，切成0.5～1平方厘米小块。 2．EDTA处理：先置入0.02％EDTA中室温置5分钟。 3．冷消化：换入0.25％胰蛋白酶中，置4℃过夜。 4．分离：取出皮肤，用血管钳或镊子把表皮与真皮层分开。 5．温消化：取出表皮单独处理，用剪刀剪成更小的块后，置入新的0.25％胰蛋白酶中，3 ...

人心房肌细胞的培养与鉴定.pdf

从活组织培养层中分离独立的细胞团.pdf

本文来自Cyagen赛业：http://www.cyagen.com.cn碱性磷酸酶（ALP）是成熟成骨细胞的标志性酶，同时成骨细胞形成的钙结节也是成骨细胞的标记物。以ALP为目标物的检测方法：1Gomori钙钴法【染色原理】ALP在pH值9.4的环境下，以镁离子作为激活剂，能够把β-甘油磷酸钠水解出磷酸，磷酸与高浓度的钙盐结合形成无色的磷酸钙，再与硝酸钴作用形成磷酸钴，经硫化胺处 ...

1.G418的配制： 取1g G418溶于1ml 1M的HEPES液中，加蒸馏水至10ml，过滤消毒，4度保存。2.细胞培养：取待测培养细胞，制备成细胞悬液，按等量接种入多孔培养板中，培养6小时左右开始加药。3.制备筛选培养基：在100ug/ml～1000ug/ml范围内确定几个梯度，比如先做个100ug/ml、400ug/ml、800ug/ml、1000ug/ml，按梯度浓度用培养基稀释G418 ...

细胞裂解分析化学论坛化学分析仪器分析分析测试色谱电泳光谱u5kLlIOT采用温和的裂解条件，不能破坏细胞内存在的所有蛋白质－蛋白质分析化学论坛~O/b:te} 相互作用，多采用非离子变性剂（NP40或Triton X-100)。每种细胞的裂解条件是不一样的，通过经验确定。 不能用高浓度的变性剂（0.2％SDS)，细胞裂解液中要加各种酶抑制剂，如商品化的cocktailer。2、如何得到比较纯 ...

1. Examine all flasks by inverted-phasemicroscopy. Cultures used for preservation should be grown free of antibiotics show no signs of microbial contamination and should be subconfluent (confluent cul ...

Remove Human ES cells from liquid nitrogen storage tank. Fill out a freeze/thaw form.Thaw cryovial by gently swirling in waterbath until only a small ice pellet remains being careful not to completely ...

based on splitting onto one plateWarm collagenase IV split media to 37 °C in a water bath.Aspirate media off of cell culture plate.Add the following amount of collagenase:0.5ml/well of 4 well plate1.0 ...

Collagenase cells for approximately 7 minutes at 37 °C (until edges of colonies are curling up).With a 5 ml pipet gently pipet and scrape colonies from plate. Add cell suspension to a 15 ml centrifug ...

Aliquoting Matrigel:Day one:Put the sterilized tip box (either 200 ml or 1000 ml tips) sterilized eppendorf tube container and appropriate pipettor in -20 °C freezer.Thaw the Matrigel bottle on ice in ...

based on splitting onto on plateWarm collagenase media to 37°C in a water bath.Aspirate media off of cell culture plate.Add the following amount of collagenase0.5ml/well of 4 well plate1.0ml /well of ...

Isolate cells and dissociate to single cell suspension (can use Gibco Cell Dissociation Buffer Accutase or Trypsin)Wash with 10% FBS/DMEM:F12For surface markers resuspend in appropriate volume of 10% ...

I. TYPES OF CELLS GROWN IN CULTURETissue culture is often a generic term that refers to both organ culture and cell culture and the terms are often used interchangeably. Cell cultures are derived from ...

DescriptionThis is a fast and easy assay to test the angiogenic/anti-angiogenic properties of molecules. As compared to other angiogenesis assays such as chicken chorioallantoic membrane assay it is c ...

8 eggs per day day 7- day 13 cut CAM wash in precooled PBS in 10 ml WASH 1 (PBS 5 mM EDTA COMPLETE) on ice cut in pieces in petri dish on ice centrifuge 4 min at 1000 rpm (table top centifuge Falcon 1 ...