lines Protocols

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Liver Toxicity Assays Protocols

This test is designed to detect irreversible toxic effects on both cell growth and survival, by the evaluation of colony-forming (CF) efficiency, in hepatoma cell lines derived from man, rat and mouse. - [Read Hepatoma Cell Cultures as In Vitro Models for Hepatotoxicity]

Category: Cell Culture Protocols: Cell Immortalization Protocols

Cultivating animal cells in the laboratory is an indispensable technique for cell biologists. However, most normal primary cell lines, while faithfully reproducing the phenotype of their tissue of origin, do not grow indefinitely in culture. After a series of population doublings (the number of which varies by species, cell type, and culture conditions) primary cells enter a state where they no longer divide. - [Read Immortalization of Cells in Culture]

Category: Immunology: Immunofluorescence Protocols

Protocol for immunofluorescence staining. Includes: Cell Lines; Frozen Sections; Paraffin Sections; Pretreatments of Tissue Sections. - [Read Immunofluorescence Staining Protocol]

Category: Viral Culture and Isolation Protocols: Viral Isolation Protocols

This protocol may be used for the infection of adherent cell lines. - [Read Infection Protocol for Adherent Cell Types]

Category: Cell Biology and Cell Culture: Cell Culture Techniques

Introduction to Animal Cell Culture. Corning. Good introduction to cell lines, cell protocols, and cell culture. - [Read Introduction to Animal Cell Culture. Corning]

Category: RNA Protocols: mRNA Extraction Protocols

Isolation of poly A+ RNA from Total RNA by Oligo(deoxythymidine)cellulose Chromatography. Total RNA is first isolated from the tissues or cells and then mRNA is isolated by PolyA+ selection using oligo(dT) cellulose. This is necessary for all tissue sources rich in RNase (and cell lines). Lazo Lab - [Read Isolation of poly A+ RNA from Total RNA by Oligo(deoxythymidine)cellulose Chromatography]

Category: Immunology: Macrophage Monocyte Protocols

Lymphoblastoid Cell Lines from Frozen Whole Blood. Rosalie Veile. Keller Lab. - [Read Lymphoblastoid Cell Lines from Frozen Whole Blood]

Category: Cell Culture Protocols: Insect Cell Culture Protocols

Insect cell cultures are now commonly used in insect physiology, developmental biology, pathology, and molecular biology. As the field has advanced from methods development to a standard procedure, so has the diversity of scientists using the technique. This paper describes methods that are effective for maintaining various insect cell lines. The procedures are differentiated between loosely or non-attached cell strains, attached cell strains, and strongly adherent cell strains. - [Read Methods for Maintaining Insect Cell Cultures]

Category: Cell Culture Protocols: Cell Culture Contamination Prevention and Removal Protocols

One of the most important, but frequently overlooked, cell culture procedures is testing cultures for microbial contamination, especially mycoplasma. It is critical for every cell culture laboratory to only use cell lines that have been carefully screened for mycoplasma. Fortunately, there is a simple fluorochrome DNA staining test that can detect both mycoplasma and virtually any other prokaryote contaminants. - [Read Mycoplasma Detection Using DNA Staining Protocol]

Category: Cell Biology and Cell Culture: Cell Electroporation Protocols

Optimization of Electroporation Transfection of DNA Protocol into Cell Lines. Eppendorf - [Read Optimization Protocol – Cell Line Electroporation PDF]

Category: Transfection Protocols

Protocol describes transfection of plasmid DNA into mammalian cell lines using electroporation, a process whereby external application of electric pulses induce cell membrane permeability. Cells in suspension and small volume cells are difficult to transfect, whereas adherent cells and large volume cells are relatively easy. Regardless of cell size or phenotype, transfection efficiency increases with a high concentration of cells in a small volume. - [Read Optimizing Electrotransfection of Mammalian Cells In Vitro Protocol]

Category: Transfection Protocols: Electroporation Sonoporation Protocols

Protocol describes transfection of plasmid DNA into mammalian cell lines using electroporation, a process whereby external application of electric pulses induce cell membrane permeability. A number of factors can affect electrotransfection efficiency. In general, cells in suspension and small volume cells are difficult to transfect, whereas adherent cells and large volume cells are relatively easy. - [Read Optimizing Electrotransfection of Mammalian Cells In Vitro Protocol]

Category: Cell Culture Protocols: Insect Cell Culture Protocols

Remove medium from culture container, rinse with PBS, expose cells to trypsin, incubate, stop trypsin activity by adding serum containing medium, dissociate cells, dilute and return to incubator. - [Read Passage of Adherent Cell Lines (subculture).]

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Preparation of metaphase chromosomes from lymphoblastoid cell lines for FISH analysis. Sanger Institue. - [Read Preparation of metaphase chromosomes from lymphoblastoid cell lines]

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

MEF feeders are prepared weekly to provide a substrate for undifferentiated embryonic stem (ES) cells. Primary MEF cells are thawed, established in culture, treated with mitomycin C to halt their proliferation so they cannot overgrow the ES cultures, and then replated onto dishes convenient for ES cell culture. This protocol can also be used to prepare feeder cells from STO fibroblast cell lines. - [Read Preparation of Mouse Embryonic Fibroblast (MEF) Feeder Plates Protocol]

Category: Molecular Imaging: Immunofluorescence Microscopy

Preparing a cell block from cell lines / cell suspensions. Cattoretti, Colombia University. - [Read Preparing a cell block from cell lines / cell suspensions]

Category: Immunohistochemistry Protocols: Blocking Protocols

Protocol for preparing a cell block from cell lines / cell suspensions. Includes: Live cell preparation; Fixed cell preparation. - [Read Preparing a Cell Block from Cell Lines Cell Suspensions Protocol]

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

This protocol is optimized for erythroid cells, other cell types may require modifications. - [Read Protocol for Stable Lines]

Category: Cell Culture Protocols: Cell Culture Contamination Prevention and Removal Protocols

Quality is important in all aspects of tissue culture since the quality of materials used i.e. media and other reagents) will affect the quality of the cultures and products derived from them. The main areas of quality control that are of concern for tissue culture are: The quality of the reagents and materials; The provenance and integrity of the cell lines; The avoidance of microbial contamination. - [Read Quality Control Considerations for Cell Culture]

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

Quality control considerations for cell culture. Includes: Provenance and Integrity of Cell Lines; Avoidance of Microbial Contamination; Environmental Monitoring; What to do in the event of contamination; - [Read Quality Control Considerations for Cell Culture]

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

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 extraction. Donis-Keller. - [Read Reactivating Cell Lines and Cell Growth for DNA Preparation]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

A powerful way to identify a mutation in the gene of interest and to test mutant plants for phenotypes that are predicted to result from loss of function of that gene is by PCR screening. Pools of insertion lines are screened using one primer corresponding to the gene of interest and one primer corresponding to the end of the insertion element. The synthesis of a product indicates the presence of an insertion in the gene of interest. - [Read Screening DNA Pools for T-DNA Insertions in Arabidopsis Genes Protocol]

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

Simple Experimental Method for Storage of Cell Lines. Hancock Lab. - [Read Storage of Cell Lines - Hancock]

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Protocol for subcloning analysis. Includes cell lines: BG01(1), TE03, TE06, UC06(1), WA01, WA07, WA09. - [Read Subcloning Analysis Protocol]

Category: Transfection Protocols: Stable Transfection Protocols

Protocol uses an autoregulatory system in which the transcriptional trans-activator tTA drives its own expression and that of a target gene. The first stage of the procedure describes how to generate stable lines of NIH-3T3 cells that express either tTA alone or tTA and the tetracycline-regulated target gene. - [Read Tetracycline as Regulator of Inducible Gene Expression Protocol]