lines Protocols

Category: Epigenetics and Methylation Protocols: Bisulfite Treatment Methylation Assay Protocol

Yang et al, NAR. 2004. ue to the heavy methylation of repetitive elements, this assay was especially useful in detecting decreases in DNA methylation, and this assay was validated by examining cell lines treated with the methylation inhibitor 5-aza-2'deox - [Read A simple method for estimating global DNA methylation using bisulfite PCR of repetitive DNA elements]

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

There are many ways to adapt cell lines to serum-free media. Five methods are presented that are designed for adapting hybridomas to a protein-free medium. These protocols may require some modifications for your particular cell line and conditions. - [Read Adapting Cells to a Serum-Free Environment Protocol]

Category: Transfection Protocols: Calcium Phosphate Transfection Protocols

This calcium phosphate transfection method works best in cell lines that are 1) highly transformed and 2) adherent (Hela, U2OS, SAOS2, AdAH, NPC-KT and obtain from 20% to 100% transfection efficiency depending on the cell line). Works well for transient experiments but precautions should be used in the design and interpretation of experiments based on the discussion below. Also works very well for generating stable cell lines. This method is quite sensitive to the amount of input plasmid. - [Read Calcium Phosphate Transfection Method]

Category: Transfection Protocols: Calcium Phosphate Transfection Protocols

This protocol,is based on the venerable method of Graham and van der Eb (1973). The procedure is used chiefly to generate stable lines of cells carrying chromosomally integrated copies of the transfected DNA. - [Read Calcium-phosphate-mediated Transfection of Cells with High-molecular-weight Genomic DNA]

Category: Reporter Gene Assays: GFP Assay Protocols

The following protocol can be used for the development of stable cell lines expressing GFP fusion proteins. Although optimal transfection procedures (e.g., calcium phosphate, electroporation, or FuGENE 6 [Roche Applied Science]) vary depending on cell type, this general transfection procedure has been successful for stable transfection of HeLa, A-431, U2OS, BHK, and HT1080 cells. - [Read Constructing and Expressing GFP Fusion Proteins]

Category: Reporter Gene Assays: Luciferase Assay Protocols

It is possible that some cell lines have lost the ability to perform RNAi or that cells derived from certain tissues do not support RNAi. This reporter assay, for RNAi in mammalian cells, can be used to establish whether the cells under study are susceptible to RNAi. - [Read Cotransfection of Luciferase Reporter Plasmids with siRNA Duplexes Protocol]

Category: Cell Biology and Cell Culture: Cell Culture Techniques: Cell Line Preservation Freezing

Cryogenic Preservation and Storage of Animal Cells. PDF.Corning Life Science. Very nice, detailed procedure for the freezing and storage of various cell lines. - [Read Cryogenic Preservation and Storage of Animal Cells - Corning Life Science. PDF.]

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

Culture conditions for various stable cell lines. - [Read Culture Conditions for Various Stable Cell Lines]

Category: Cell Biology and Cell Culture: Cell Line Culture Conditions

Culture of Human Prostatic Carcinoma Cell Lines, Growing and splitting the cells, Preparation of frozen stocks in liquid nitrogen, How to bring frozen cells back into culture, Concentrations of antibiotics for the selection of stable transfectants. LNCa - [Read Culture of Human Prostatic Carcinoma Cell Lines]

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Culture conditions have been established for a second blastocyst-derived cell line, trophoblast stem (TS) cells, in addition to embryonic stem (ES) cells. This protocol describes a method for culturing TS cell lines. These cells can then be used to study trophoblast differentiation and placental function. - [Read Culturing Trophoblast Stem (TS) Cell Lines Protocol]

Category: Signalling Signal Transduction Protocols: Cytokine Protocols

Cytokine-Induced Proliferation of Indicator Cell Lines, TNF-α-Induced Killing of L929 Cell Line, IFN-γ Protection from Viral Infection of Cell Lines. eBiosciences. - [Read Cytokine Bioassays Protocols]

Category: Signalling Signal Transduction Protocols: Cytokine Protocols

Antibody Neutralization of TNF-α-Induced Killing of L929 Cell Line, IFN-γ-Protection from Viral Infection of L929 and A549 Cell Lines, TNF-α-Induced Killing of L929 Cell Line. eBiosciences. - [Read Cytokine Neutralization, In Vitro Using Antibodies]

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

The starting material for de novo isolation of stem cell lines can be either normal 3.5-days post coitum (dpc) expanded blastocysts or "delayed" blastocysts. Delayed blastocysts are usually collected 4-6 days after ovariectomy. For both groups of blastocysts, tissue culture procedures are similar. The only difference is the timing of the first disaggregation, because delayed blastocysts will initially grow more slowly. - [Read De Novo Isolation of Embryonic Stem (ES) Cell Lines from Blastocysts Protocol]

Category: Stem Cell Protocols

This protocol decribes derivation of TS cell lines from 3.5-days post coitum (dpc) mouse blastocysts. The procedure is similar to the derivation of embryonic stem (ES) cell lines. However, the success rate is considerably higher, and less expertise is required to recognize pluripotent TS cell colonies. - [Read Derivation of Trophoblast Stem (TS) Cell Lines from Blastocysts Protocol]

Category: Cell Culture Protocols: Insect Cell Culture Protocols

Using this method, insect cells should be adapted to HyQ SFX-Insect in 4-8 passages. However, there are some cell lines and clones that are more sensitive to the physiochemical and nutritional changes, and in some cases it may take longer than 8 passages for the adaptation. - [Read Direct Adaptation of Insect Cells to HyQ© SFX-Insect™ Medium]

Category: Transfection Protocols: Stable Transfection Protocols

Pulsed electrical fields can be used to introduce DNA into a wide variety of animal cells. Electroporation works well with cell lines that are refractive to other techniques, such as calcium phosphate-DNA coprecipitation. But, as with other transfection methods, the optimal conditions for electroporating DNA into untested cell lines must be determined experimentally. - [Read DNA Transfection by Electroporation]

Category: Drosophila Protocols: Drosophila Culture Media Protocols

Protocol for Drosophila growth conditions. Includes: MAINTENANCE; Semi-adherent cell lines; Adherent cell lines; SL2, S2C, S2*; DL2, DL1, S2R+, Kc167, Clone 8, S3; BG2. - [Read Drosophila Growth Conditions Protocol]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture

Electroporation and isolation of ES cell lines Mouse. BayGenomics. - [Read Electroporation and isolation of ES cell lines Mouse]

Category: Transfection Protocols: Electroporation Sonoporation Protocols

Protocol for electroporation of Burkitt's Lymphoma cell lines. - [Read Electroporation of Burkitt's Lymphoma Cell Lines Protocol]

Category: Cell Biology and Cell Culture: Cell Electroporation Protocols

Electroporation of Cell Lines With DNA. Electroporation for the efficient transfection of mammalian cells with DNA. Chu et al. Cellular Immunology Oxford - [Read Electroporation of Cell Lines With DNA]

Category: Cell Biology and Cell Culture: Cell Electroporation Protocols

The protocol uses a Bio-Rad Gene Pulser II apparatus to electroporate DNA into human cell lines. Dr. Frank, Arthritis and Immunology Program, Oklahoma City Medical Research Foundation - [Read Electroporation of Eukaryotic Cells]

Category: Cell Biology and Cell Culture: Cell Culture Techniques: Cell Line Preservation Freezing

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. Kitto Lab Texas. - [Read Freezing and Thawing of Mammalian Cell Lines]

Category: Cell Culture Protocols: Cell Storing and Cell Freezing Protocols

For long term storage of myeloma cells, hybridoma cells, T cells, and other mammalian cell lines in liquid nitrogen, and restoring them in culture. - [Read Freezing and Thawing of Mammalian Cell Lines Protocol]

Category: Cell Biology and Cell Culture: Cell Culture Techniques: Cell Line Preservation Freezing

Freezing media, DMSO, DMSO Storage of cell lines protocol and methods. Suprya Jayadev - [Read Freezing Cells]

Category: Cell Biology and Cell Culture: Cell Culture Techniques

ECACC Handbook of Cell Culture. Fundamental Techniques in Cell Culture: A Labratory Handbook. Sigma Aldrich. Excellent Guide to Cell Culture, aseptic technique, storage of cell lines, subculture of cells. - [Read Handbook of Cell Culture. - Sigma Aldrich.]