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

美国实验室wetern方法

WESTERN BLOT PROTOCOLIn a Western blot, proteins that are separated on polyacrylamide gels on the basis of size are transferred to a membrane for dete

ELISA

Gangliosides ELISA protocol (Contributed by pingsunjim)This protocol can be used for detection of gangliosides.Specific antibodies to gangliosides are

细胞培养——细胞保藏

Working Cell Bank (Contributed by Nanci Donacki)Provides detailed protocol for establishing a working cell bank    Master Cell Bank (Contributed by Na

Human-Embryonic-Stem-(ES)-Cell-Protocols——Thawing-Human-ES-cells

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

Human-Embryonic-Stem-(ES)-Cell-Protocols——Freezing-Human-ES-Cells

 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 f

Western-blotting样品准备-(二)

Sodium orthovanadate preparationAll steps to be performed in a fume hood.          a. Prepare a 100 mM solution in double distilled water.          b.

Isolation-of-cell-nuclei-for-the-application-in-the-cellfree-system

Characteristics of the procedurePreparation of isolated nuclei - procedurePreparation of radioactive labeled nucleiMaterial Characteristics of the pro

A-novel-method-of-growing-fungi-for-DNA-extraction

Preparation of fungi for DNA extraction typically involves growing cultures in liquid culture in Erlenmeyer flasks, Roux bottles or even microfuge tub

The-Effects-of-NiCl2on-Spicule-Formation

The Effects of NiCl2on Spicule FormationJessica Ann Billet, Franklin and Marshall, Class of 2000Background and ObjectiveSea urchins exhibit radial hol

Assay-of-Tyrosine-Kinases-Using-Synthetic-Peptides

实验概要        Small synthetic peptide substrates are especially well suited for applications such as assays of tyrosine kinases in permeabilized cel

Preparing-chemically-competent-cells

MaterialsPlate of cells to be made competentTSS bufferLB mediaIceGlassware & EquipmentFalcon tubes500μl Eppendorf tubes, on ice200ml conical flask200μ

Fibroblast-Cell-Systems3

Seeding After cells are thawed:NOTE: Do not dispense the entire contents of the cryovial into one T-25 flask!!Remove the cap, being careful not to tou

Measurement-of-Cell-Adhesion-Under-Static-Conditions

Many different molecules have been described to promote cell adhesion including several cell surface carbohydrate-binding proteins. Measuring cell adh

酵母转化

·         Yeast Transformation (Gietz Lab)LiAc/SS-DNA/PEG Transformation·         Yeast Transformation (Breeden Lab)LiAc method·         Large-Scale Y

Using-a-Counting-Chamber

Using a Counting ChamberFor microbiology, cell culture, and many applications that require use of suspensions of cells it is necessary to determine ce

HOW-TO-USE-THE-COULTER-COUNTER-TO-COUNT-CELLS

1) Turn on the counter by pulling out the on/off button. You need to do this at least 10 min before use to obtain sufficient vacuum.Usually put 0.2 ml

Immunofluorescent-Staining-of-Drosophila-Larval-Brain-Tissue

实验概要The Drosophila larval brain is a well-established model for investigating the role of stem cells in development. Neuroblasts (neural stem cells)

Automated-Genomic-DNA-Extraction

实验概要This section  provides a general protocol for automated isolation of genomic DNA from  10-20 µl blood samples in a 96-well format using the Charge

Detection-of-MicroRNA-Heterogeneity-in-Single-Cells-Using-an-Automated

Introduction  MicroRNA  (miRNAs) are short (18–24 nucleotides), non-coding RNAs that regulate  gene expression by both disrupting messenger RNA (mRNA

High-Molecular-Weight-Yeast-Liquid-DNA-Preparation

Purpose:To isolate intact, high molecular weight DNA from yeast cells for subcloning and rare cutting restriction enzyme analysis. One can expect a yi

E.Z.N.A.®-Plasmid-Maxi-Kit-vacuum-Protocol

实验概要This Protocol is designed to isolate 500-1200 ug of high Copy-Number plasmids or 50-400 ug of low Copy-Number Plasmids from 200 ml overnight c

MITOMYCIN-C-TREATMENT-OF-PMEFs

Cultures to be treated should be sub confluent ie actively growing.1. Add 1/20 volume Mitomycin C (200 ug/ml 10 ug/ml), to culture and incubate at 37

胞外基质

ECM Cell Attachment Assay (LTI)Cell Adherence Inhibition Assay (LTI)General protocol--Either monoclonal antibody or RGD peptide is added along with th

Human-B-cell-isolation-and-culture

实验概要This protocol provides a general protocol for human B cell isolation and culture.实验步骤B cell isolation1. Donor blood was obtained with informed con

实验室自动化与筛选协会2013亚洲会展Tecan展商技术论坛

  Tecan专场时间:2013年6月6日下午13:45-14:30   Tecan Exhibitor Tutorial:Day 1 ( 6 June,2012) 13:45-14:30   Tecan专场地点:上海金茂君悦大酒店宴宾厅 II   Location: Drawing Room

Heterogeneity-of-SingleCell-Gene-Expression-Across-Phenotypically(一)

Introduction  Multi-cellular  populations are fundamentally driven by the collective properties of  individual cells. However, our understanding of ge

MN-in-Human-Lymphocytes-(method-description)

MN in Human Lymphocytes (method description)Lymphocyte isolation    Lymphocytes were isolated using Ficoll-Paque density gradients. Blood was diluted

Streptomyces:Protocols/Conjugation

Intergeneric Conjugation and OverlayDescription Transfer of plasmid/cosmid DNA from a host strain, e.g. E.coli ET12567 [pUZ8002], to the recipient str

Miniprep/Qiagen-kit

MaterialsFor purifying plasmid DNA from Escherichia coli cells, the Qiagen Spin Miniprep Kit produces quite reliable results.Do not autoclave solution

Automated-Extraction--Normalized-DNA-Buccal-Kit

实验概要This section  provides a general protocol for automated isolation of genomic DNA from  human buccal cell swabs in a 96-well format using the Charg