FreezingandThawingofMammalianCellLines
For long term storage of myeloma cells, hybridoma cells, T cells, and other mammalian cell lines in liquid nitrogen, and restoring them in culture.FreezingPreparationCells are to be frozen in liquid nitrogen, so make sure your canisters are relatively full of nitrogen and you have room. Cells should be healthy (>90% viability) and growing in log phase. You will also require sterile 1 mL cryo-vials; they have a scr......阅读全文
Growth,－Maintenance－and－Transfection－of－Suspension－Adapted－293EBNA－cells
ProcedureI. INTRODUCTIONThe 293 EBNA cell line is established from primary embryonal human kidney cells transformed with sheared human adenovirus type
Freezing－Cells
1) Keep prepared solutions on ice.2) Determine total cell count of cells to be frozen. (e.g. 1 X 108 )3) Determine number of vials to be frozen. (e.g.
巨噬细胞和单核白细胞
· Lymphocyte Transformation (Donis-Keller lab)Lymphocytes are transformed to establish cell lines. Mononuclear cells (lymphocytes) from antico
stem－cell－culture－protocol
实验概要stem cell culture protocol主要试剂cell culture supplies and reagentssEnvironment: cell culture requires a sterile environment, so it needs a separat
哺乳动物细胞(mammalian－cell)总RNA的分离1
哺乳动物细胞总ＲＮＡ的分离细胞的裂解提取步骤注意事项这一从培养的单层哺乳动物细胞中分离RNA的实验程序系为Favaloro 等（1980）所介绍程序的改良。该程序同样适用于从悬浮培养的哺乳动物细胞中或从易于分散成单个细胞的哺乳动物组织中分离 RNA。但不适用于从固体组织中提取RNA，因为用这
哺乳动物细胞(mammalian－cell)总RNA的分离2
3）于0℃以5000g离心10分钟沉淀RNA，弃上清，用含0.1mol/L 乙酸钠（pH5.2）的70％乙酸洗涤沉淀，用自动微量移液器尽可能将乙醇吸尽， 随后于室温放置几分钟，晾干沉淀。切勿抽干沉淀，因为干的核酸沉淀很难溶解。4）每个直径为90mm的培养皿或每107细胞加 200μl 50mmol/
Glycosylation－of－Antibody－Therapeutics:－Optimisation－for－Purpose
Recombinant antibody therapeutics represent a significant success story in terms of clinical benefit delivered and revenue (profit) generated with
病毒冷冻保藏技术
实验概要Snap freezing, or flash freezing, is the process by which samples are lowered to temperatures below -70°C very rapidly using dry ice or liquid
Examination－of－a－Mammalian－Blood－Smear
Examination of a Mammalian Blood SmearThe distribution and appearance of the formed elements of blood can tell a great deal about the condition of the
Culturing－Mouse－Embryonic－Fibroblasts
MaterialsTrypsin (Gibco 25200-023)3T3 Medium: 500 mL DME (Invitrogen) + 50 mL FBS (Hyclone) + 5 mL 100x Pen/Strep2x Freezing Medium: 3T3 Medium + 20%
Routine－Splitting－and－freezing－of－cells
1. Grow cells to subconfluence in a flask.2. Harvest as per normal and count.3. Spin down 5min 1.2K in benchtop. Resuspend at 1.0 X 106/ml in 10% DMSO
Freezing－cells－in－liquid－nitrogen
Take off MediaTrypsinate with 1ml x2 Dulbecco A trypsinAdd 7ml MediaPipette up and down to distribute cells throughout media (i.e. not clumped togethe
Comparison－of－Enzymatic－and－NonEnzymatic－Means3
MTT Assay on Reattached CellsAs seen in Fig. 2 , the proportion of viable MSC that re-attached was significantly higher (p = 0.0004) upon dissociati
Transfection－of－Mammalian－Cells－Using－Lipofectamine
Materials: LipofectamineBasal Medium containing 10% fetal bovine serum, 1% glutamine, 1% aaBasal Medium containing 1% glutamineBasal Medium cont
Serum－Thawing－－Heat－Inactivation
How to thaw serum:Serum that is stored at -10º C to -40º C is stable for extended periods of time. It is neither necessary or desirable to store serum
Serum－Thawing－－Heat－Inactivation
Serum Thawing & Heat Inactivation(Chris Cohick from JRH Biosciences catalogue)How to thaw serum:Serum that is stored at -10º C to -40º C is stable for
Thawing－and－Plating－Cryopreserved－Hepatocytes
实验概要This protocol covers thawing and prep of cryopreserved hepatocytes for applications such as metabolic stability, intrinsic clearance, enzyme in
Cryopreservation－of－cell－cultures
1. Examine all flasks by inverted-phasemicroscopy. Cultures used for preservation should be grown free of antibiotics, show no signs of microbial cont
Comparison－of－Enzymatic－and－NonEnzymatic－Means2
MTT Assay on Reattached CellsMSC were seeded in 12-well cell culture dishes with 5.0 × 104 cells per well (≈4.8 cm2). After 5 to 6 days of culture, co
Production－of－Antibody－Fragments－in－Arabidopsis－Seeds
Plants offer a number of attractive benefits over conventional mammalian or bacterial cell culture systems for the production of valuable pharmace
Thawing－Cells－(Schreibers－protocol)
Thaw vial quickly in 37癈 water. Caution - vial can explode.Transfer cells to sterile, 15 mL centrifuge tube.Add 50 祃 warm FBS (fetal bovine serum, hea
抗生素溶液配制
Working concentrations and stock solutionsPreparation of stock solutionsAmpicillinCarbenicillinGE antibiotic mixTetracyclineChloramphenicol Supplement
Genomic－DNA－Extraction－－PureLink™
实验概要The PureLink™ Genomic DNA Purification Kit allows rapid and efficient purification of genomic DNA. The kit is designed to efficiently isolate g
Steps－in－the－Glycosylation－of－Mammalian－Nlinked－Oligosaccarides
The biosynthesis pathway of N-glycans is a costly system with respect to the number of enzymes that are involved in the synthesis and trimming of N-gl
Methods－for－the－Measurement－of－a－Bacterial－Enzyme－Activity－in－Cell－Lysates2
Measurement of ACTase activityNuclear magnetic resonance spect roscopy (NMR)The unique potential of NMR spectroscopy for monitoring simultaneously the
Infection－with－retroviruses
It is well established with avian retroviruses that cells are most efficiently infected just after they are trypsinized. Trypsinization does two thing
Thawing－－Incubating－Human－－Animal－Liver－Microsomes
实验概要BackgroundThe liver is the major organ for metabolism of endogenous substrates as well as exogenous drugs. There are several in vitro tools avai
Culturing－Human－Neural－Stem－Cells
实验概要Neural stem cells (NSC) are valuable resources because of their ability to differentiate into neurons and glial cells with applications in neur
哺乳动物RNAi技术－Mammalian－RNA－Interference
Mammalian RNA InterferenceThomas TuschlLaboratory for RNA Molecular BiologyThe Rockefeller University, New York Excerpted from RNAi: A Guide To Gene S
Culture－of－BEND－Cells－(Bovine－Endometrial－Cells)
Culture of BEND Cells (Bovine Endometrial Cells)Charles E. Krininger, III and Peter J. Hansen Dept. of Animal Sciences, University of FloridaThis prot