ResuscitationofFrozenCellLines
AimMany cultures obtained from a culture collection, such as ECACC, will arrive frozen and in order to use them the cells must be thawed and put into culture. It is vital to thaw cells correctly in order to maintain the viability of the culture and enable the culture to recover more quickly. Some cryoprotectants, such as DMSO (Prod. No. D2650), are toxic above 4oC therefore it is essential that cultures are thaw......閱讀全文
Resuscitation－of－Frozen－Cell－Lines
AimMany cultures obtained from a culture collection, such as ECACC, will arrive frozen and in order to use them the cells must be thawed and put into
Method:－Lymphoblastoid－Cell－Lines－from－Frozen－Whole－Blood
Method: Lymphoblastoid Cell Lines from Frozen Whole BloodMay 31, 1990Rosalie VeilePurpose:Blood Samples can be stored frozen as a backup in case an LC
Cryopreservation－of－Cell－Lines
AimThe protocol below describes the use of passive methods involving an electric -80oC freezer for the cryopreservation of cell cultures. ECACC routin
Subculture－of－Suspension－Cell－Lines
AimIn general terms cultures derived from blood (e.g. lymphocytes) grow in suspension. Cells may grow as single cells or in clumps (e.g. EBV transform
Subculture－of－Adherent－Cell－Lines
AimAdherent cell lines will grow in vitro until they have covered the surface area available or the medium is depleted of nutrients. At this point the
Method:－Maintaining－Lymphoblastoid－Cell－Lines
Method: Maintaining Lymphoblastoid Cell LinesJune 10, 1990Rosalie VeilePurpose:To grow lymphoblastoid cells for permanent storage and for DNA extracti
Freezing－and－Thawing－of－Mammalian－Cell－Lines
For long term storage of myeloma cells, hybridoma cells, T cells, and other mammalian cell lines in liquid nitrogen, and restoring them in culture.Fre
Method:－Reactivating－Cell－Lines－and－Cell－Growth－for－DNA－Preparation
Purpose:Cell lines are reactivated and grown to a count of 1 x 108 cells. The cells are pelleted and stored frozen at -80 degrees C prior to DNA extra
Subculture－of－SemiAdherent－Cell－Lines
AimSome cultures grow as a mixed population (e.g. B95-8 - marmoset) where a proportion of cells do not attach to the tissue culture flask and remain i
Generating－stable－cell－lines－in－HEK293
Generating stable cell lines in HEK293Prior to transfection, it is recommended that you linearize your pcDNA gene construct. Linearizing will decrease
Generating－stable－cell－lines－in－HEK293
Generating stable cell lines in HEK293Prior to transfection, it is recommended that you linearize your pcDNA gene construct. Linearizing will decrease
Method:－Preparation－of－Lymphoblastoid－Cell－Lines－for－Long－Term－Storage
Method: Preparation of Lymphoblastoid Cell Lines for Long Term StorageMay 30, 1990Rosalie VeilePurpose:To store cell lines in a form that will insure
Method:－Logging－in－Specimens－and－Record－Keeping
Method: Logging in Specimens and Record KeepingJune 10, 1990Rosalie VeilePurpose:To keep a written and computerized record of all cell lines, the date
巨噬細胞和單核白細胞
·?????????Lymphocyte Transformation?(Donis-Keller lab)Lymphocytes are transformed to establish cell lines. Mononuclear cells (lymphocytes) from antico
細胞培養——細胞保藏
Working Cell Bank?(Contributed by?Nanci Donacki)Provides detailed protocol for establishing a working cell bank????Master Cell Bank?(Contributed by?Na
細胞培養常規操作
常規操作(主要內容如下)·?????????Aseptic Technique·?????????Culture Vessels·?????????Cell Counting·?????????Primary Culture·?????????Maintenance of Cell Line?·??
Freezing－and－Thawing－of－MEFs
Author:?Shalini Jain and Hariom YadavAffiliation:?Animal Biochemistry Division, National Dairy Research Institute, Karnal-132001, Haryana, IndiaDate A
IHC－frozen－sections...
實驗概要The method provides a guideline procedure and tips for staining of frozen sections.實驗步驟Frozen sections: Once mounted on APES coated slides, frozen
Immunohistochemistry－Protocol－for－Frozen－Sections
實驗概要The ?following is a general procedure guide for preparation and staining of ?acetone-fixed frozen tissues using a purified, unconjugated primary ?
Immunohistochemistry－Protocol－for－Frozen－Sections
實驗概要The ?following is a general procedure guide for preparation and staining of ?acetone-fixed frozen tissues using a purified, unconjugated primary ?
Preparation－and－Staining－of－Frozen－Tissue－Sections
I. Preparation of Frozen Sections for SectioningMaterials neeed:2-methylbutane (isopentane)Liquid NitrogenDry icePeel-Away?/sup> base moldsFrozen tiss
Extraction－of－RNA－from－Frozen－Sections
RNA Extraction from Frozen Tissue Sections?Tissue Handling:?Note that all unfixed human tissue should be handled as BioSafety Level 2 materials (wear
Apoptosis－TUNEL－Assay－(frozen－sections)
Protocol for Frozen Sections:Warm 150ml 4% Paraformaldehyde/1x PBS to RT. Fix slides in it, 20 min., RT.1x PBS rinse, 2 times.1x PBS, 30 min., RT. Beg
The－Dos－and－Donts－of－Cell－Culture
Given below are a few of the essential "do’s and don'ts" of cell culture. Some of these are mandatory e.g. use of personal protective equipment (P
DNA－Extraction－from－Frozen－Tissue－Sections
Tissue collection, storage, microdissection, sectioning: See separate protocol.Tissue handling: Note that all fresh tissue should be handled as BioSaf
Fungal－Midi－DNA－Kit－Protocol－for－Fresh/Frozen－Specimens
實驗概要This ?protocol is suitable for most fresh or frozen tissue samples allowing ?more efficient recovery of DNA. However, due to the tremendous variat
Growing－feederindependent－embryonic－stem－cells§
We use feeder-independent ES cell lines derived from the 129/Ola strain of mice (Nichols et al., Development 110, p.1341, 1990). These cells are easy
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
Double－immunofluore...
實驗概要We provide a protocol for immunofluoresent double staining incubating the antibodies together.In order to be able to examine the co-distributi
ES－Cell－Culture－and－Manipulation
MediaHigh glucose DMEM (-pyruvate, -glutamine)20% Heat-inactivated Fetal calf serum (can vary by cell type, be sure!!)1X l-glutamine1X Penicillin/stre