ProtocolforCellFusion
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 clinical centrifuge (in 50 ml tubes) for two minutes. The media should be removed and the cells resuspended in fresh media. This ensures that only the healthiest cells will fall to the bottom of the tube. Centrifugation at......阅读全文
Chemotaxis－Assay趋化性实验
Springer Lab，The CBR Institute for Biomedical Research, Inc. Department of Pathology Harvard Medical Schoolhttp://cbr.med.harvard.edu/investigators/sp
Blackburn:Yeast－Colony－PCR
OverviewThis is a quick and easy yeast colony PCR protocol that does not require zymolyase step.Updated Protocol: Blackburn Lab: Quick and Easy Yeast
Isolation－of－microRNA－(miRNA)
实验概要 This protocol utilizes the powerful guanidine isothiocyanate–phenol:chloroform extraction method which allows the rapid isolation of t
293fectin™－Transfection
实验概要293fectin™ is a proprietary, cationic lipid-based formulation for transfection of DNA into eukaryotic cells. 293fectin™ is optimized for transfe
Cell－and－tissue－lysis－hub
This page should point you to the many different general and lab-specific protocols describing tissue and cell lysis and serve as forum for comparison
Combined－Flow－Cytometric－Measurement－of－Two－CellSurface－Antigens2
DNA and RNA Staining6. Stain cells with 7-AAD: i. Resuspend the cells from Step 5 in 0.5 mL of NASS containing 10 µg/mL of 7-AAD. Incubatefor 20 min a
同位素法测定底物磷酸化活性方法
实验概要Ideally, one would like to be able to directly phosphorylate substrates in an intact cell. This could potentially be performed by introducing AT
CO2恒温摇床解决人胚肾－293－(HEK293)－细胞结团问题（一）
人胚肾 293 (HEK293) 细胞在重组蛋白表达中是最常见的宿主细胞。 这类细胞能够表达大量的膜蛋白，如 G 蛋白偶联受体 (GPCR) ，是无法在最常见的生物制药生产宿主，如：中国仓鼠卵巢 (CHO) 细胞中作表达。 HEK293 虽然是蛋白表达的极好宿主，然而 HEK293 细胞
Immunodetection－of－cyclin－D1－and－D2/D3－using－flow－cytometry
DescriptionThis protocol is for use with the D cyclins and employs 488 nm argon laser excitation of propidium iodide and 630 nm NeNe or diode laser ex
CHO－Centrosome－Prep
CHO Centrosome Prep:Arshad Desai4/94Cells:We grow our CHOs with MEM[[alpha]] (without nucleosides) + 10% Bovine Calf Serum and penn/strep/glutamine. F
Basic－Method－for－Indirect－Immunofluorescence－Labeling
Basic Method for Indirect Immunofluorescence LabelingBackgroundThis is the method for indirect immunofluorescence labeling; that is, the antibodies do
Protein－Expression－and－Purification－Protocol
Step 1: Transform appropriate DNA plasmid into BL21(DE3) E. coli cells. These cells must be competent. (Protocol for how to make competent cells.)a) T
Western－blotting样品准备－（一）
实验概要Preparation of lysis buffers, protease and phosphatase inhibitors, lysate from cell culture, lysate from tissues, protein concentration, samples
Xenofree－Culture－of－Neural－Stem－Cells
实验概要Neural stem cells (NSCs) derived from human embryonic stem cells (hESCs) have the potential to help provide understanding for human neurogenesis
ChIP－using－plant－samples－–－Arabidopsis
实验概要This protocol describes how chromatin is prepared from Arabidopsis, which can subsequently be used for chromatin immunoprecipitation (ChIP). T
侧群干细胞（sp细胞）的分离与分选
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； DNase I 100*; PI：50ug/ml; Hoechst33342: 1000ug/ml（all from Sigma） 计数板，冰盒；各种离心管，无菌流式管（BD），400滤网（BD
Streptomyces:Protocols/MiniMaxi－Prep
Small Scale Plasmid Isolation (Mini / Maxi Prep)Description A mini prep / maxi prep is used to isolate plasmid or cosmid DNA from bacteria, normally E
夹心ELISA2
General Protocol for the Sandwich ELISA method:Before the assay, both antibody preparations should be purified and one must be labeled.For most applic
Tissue－Culture－Methods3
REFERENCES:R. Ian Freshney, Culture of Animal cells: A manual of basic techniques, Wiley-Liss, 1987.VI. TISSUE CULTURE PROCEDURESEach student should m
Optimized－Method－for－the－Preparation－of－Rodent－Testicular－Cells1
Homogeneity of cell populations is a prerequisite for the analysis of biochemical and molecular events during male gamete differentiation. Given the c
CarbohydrateSpecific－Adhesion－of－Intact－Cells－to－Resolved－Glycolipids－on－T
Carbohydrate-Specific Adhesion of Intact Cells to Resolved Glycolipids on TLC PlatesRonald L. Schnaar~Professor, Johns Hopkins University Medical
DNA转化
DNA转化Chemical Transformation· Transformation of Competent Cells (RbCl2 Method) (Goldberg Lab)Very nice protocol for E. Coli transformation inc
Activation－and－Expansion－of－Human－Treg－Cells
实验概要This protocol is intended for activation and expansion of human Treg cells isolated with the Dynal® CD4 CD25 Treg Kit (Cat. no. 113.23D). The exp
Preparation－of－Plasmid－DNA－by－Alkaline－Lysis－with－SDS:－Maxipreparation
实验概要Plasmid DNA is isolated from large-scale (500 ml) bacterial cultures by treatment with alkali and SDS.主要试剂Buffers and SolutionsAlkaline lysis solu
核蛋白基因组指纹技术
Fine Mapping of Genomic Targets of Nuclear Proteins in Cultured CellsAchim Breiling and Valerio OrlandoDulbecco Telethon Institute, Institute of Genet
免疫荧光
Immunofluorescence Technique (Spector Lab)protocol for immunofluorescence on cells Immunofluorescence Protocol (Walter Steffen)Methanol fixationForma
mpulsive－Pressurization－of－Neuronal－Cells－for－Traumatic－Brain－Injury－Study
实验概要A novel impulsive cell pressurization experiment has been developed using a Kolsky bar device to investigate blast-induced traumatic brain injur
Dual－ELISPOT
实验概要We provide a protocol using an FITC-conjugated primary antibody and a biotinylated primary antibody. Those are in turn recognized by anti-FITC H
Example－Tissue－Bank－Protocol1
BackgroundSummarize the purpose and objectives of the tissue bank in this section.2 Eligibility to Participate in the Research Tiss
Performing－a－hunt－by－interaction－mating
AbstractWhen more than one bait will be used to screen a single library, significant time and resources can be saved by performing the interactor hunt