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CALCIUM PHOSPATE TRANSFECTION OF HUMAN CELLS - http://omrf.ouhsc.edu/~frank/CAPO4TRANSFECT.html

Category: Cell Biology and Cell Culture: Cell Transfection Protocol

Simple Transfection using Calcium chloride and phosphate.Dr. Frank. Arthritis and Immunology Program, Oklahoma City. Medical Research Foundation. - [Read CALCIUM PHOSPATE TRANSFECTION OF HUMAN CELLS]

ChIP on ChIP Protocol PDF - http://genomecenter.ucdavis.edu/farnham/farnham/protocols/MOJO%20Dec%202003%20ChIP-chip%20protocol.pdf

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

Method to identify genomic targets of DNA-binding factors. Chromatin immunoprecipitation (ChIP) assay on high-throughput microarray based methods for discovering genomic regions occupied by human DNA-binding proteins. Oberley and Farnham. - [Read ChIP on ChIP Protocol PDF]

Human Hepatocyte Culture Protocol - http://www.montp.inserm.fr/u632/PDF/Pichard2006%20MethMolBiol.pdf

Category: Cell Biology and Cell Culture: Hepatocyte Cell Culture

Primary culture of hepatocytes is an in vitro model widely used to ... protocols for cytochromes P450IIIA in vivo and in primary cultures of animal and ...Human Hepatocyte Culture Protocol - [Read Human Hepatocyte Culture Protocol]

Electroporation of Eukaryotic Cells - http://omrf.ouhsc.edu/~frank/EPOREUK.html

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]

Calcium-phosphate-mediated Transfection of Eukaryotic Cells with Plasmid DNAs - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3871

Category: Transfection Protocols: Calcium Phosphate Transfection Protocols

Calcium phosphate forms an insoluble precipitate with DNA, which attaches to the cell surface and is taken into the cells by endocytosis. The protocol is easily adapted for use with other types of cells, both adherent and nonadherent. This protocol is a modified version of a method published by Jordan et al. (1996) who rigorously optimized calcium-phosphate-based transfection methods for Chinese hamster ovary cells and the 293 line of human embryonic kidney cells. - [Read Calcium-phosphate-mediated Transfection of Eukaryotic Cells with Plasmid DNAs]

Transient Transfection Into 293T Cells Protocol - http://www.cbrinstitute.org/labs/springer/protocols/jun_293T.html

Category: Transfection Protocols: Transient Transfection Protocols

Transient transfection into 293T cells is a convenient way to overexpress and obtain both cellular and extracellular (secreted or membrane) proteins. 293 is a human renal epithelial cell line which is transformed by adenovirus E1A gene product. 293T is a derivative which also express SV40 large T antigen, allowing episomal replication of plasmids containing the SV40 origin and early promoter region. They (both) have the unusual property of being highly transfectable. - [Read Transient Transfection Into 293T Cells Protocol]

A high yield affinity purification method for specific RNA-binding proteins. - http://www.pubmedcentral.gov/pagerender.fcgi?tool=pmcentrez&pageindex=1&artid=330202

Category: RNA Protocols: RNA-binding Protein Purification

A high yield affinity purification method for specific RNA-binding proteins: isolation of the iron regulatory factor from human placenta. Neupert 1990. - [Read A high yield affinity purification method for specific RNA-binding proteins.]

2D Gel Electrophoresis for Human Plasma Proteome - http://web.archive.org/web/20031115200452/http://iprotocol.mit.edu/protocol/362.htm

Category: Proteomic Protocols: 2-D Gel Electrophoresis

2D Gel Electrophoresis for Human Plasma Proteome. Lee Lab. - [Read 2D Gel Electrophoresis for Human Plasma Proteome]

Restraint and Handling of Animals Guide - http://www.ahc.umn.edu/rar/handling.html

Category: Animal Research Techniques

Handling Methods for animals: mice, rats, hamsters, rabbits, cats, dogs, non-human primates, swine, goats, sheep, calves, pigs. Restraint devices, drug administration. - [Read Restraint and Handling of Animals Guide]

Human T Cell Activation Protocol - http://www.ebioscience.com/ebioscience/appls/AC145.htm#human

Category: Immunology: T Cell Protocols

Protocol for the Stimulation of human peripheral blood mononuclear cells with anti-human CD3 monoclonal antibody; MTT assay for detection of cellular proliferation. Human PBMCs can be activated in vitro by soluble anti-human CD3 antibodies. Performed titration studies with these antibodies and established the following protocol for stimulation of PBMC. - [Read Human T Cell Activation Protocol]

Fluorescence In Situ Hybridization (FISH) on Human Chromosomes and Interphase Nuclei Protocol - http://users.ugent.be/~fspelema/neubla/protocol/fish.htm

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Protocol for fluorescence in situ hybridization (FISH) on human chromosomes and interphase nuclei. Includes: Plasmid probes; COSMID-, YAC- and LIBRARY PROBES; Probe detection. - [Read Fluorescence In Situ Hybridization (FISH) on Human Chromosomes and Interphase Nuclei Protocol]

Culture of Human Embryonic Stem Cells (hESC) - http://stemcells.nih.gov/research/NIHresearch/scunit/culture.asp

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

Culture of Human Embryonic Stem Cells (hESC). one simple protocol outlined. NIH Stem Cell Unit - [Read Culture of Human Embryonic Stem Cells (hESC)]

Growth of Primary Cell Culture and Viral Vector Handling Protocols - http://www.stanford.edu/dept/EHS/prod/researchlab/bio/docs/Palmer_Bio_protocol.pdf

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

The procedures involve the isolation and growth of primary cell cultures from rodent and human tissue as well as the use of viral vectors for the introduction and expression of mammalian genes in cells in culture and in live rodents. - [Read Growth of Primary Cell Culture and Viral Vector Handling Protocols]

Culture and Characterization of Human Embryonic Stem Cells. PDF - http://www.liebertonline.com/doi/pdf/10.1089/scd.2004.13.325

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: ES Cell Growth Media and Requirements

Culture and Characterization of Human Embryonic Stem Cells. DRAPER et al., STEM CELLS AND DEVELOPMENT 13:325Ã336 (2004) - [Read Culture and Characterization of Human Embryonic Stem Cells. PDF]

Isolation and Growth of Primary Cell Cultures from Mouse Protocol - http://www.stanford.edu/dept/EHS/prod/researchlab/bio/docs/Palmer_Bio_protocol.pdf

Category: Mouse Protocols: Mouse Cell Culture Protocols

The procedures involve the isolation and growth of primary cell cultures from rodent and human tissue as well as the use of viral vectors for the introduction and expression of mammalian genes in cells in culture and in live rodents. - [Read Isolation and Growth of Primary Cell Cultures from Mouse Protocol]

Culture of Human Embryonic Stem Cells (hESC) Protocol - http://stemcells.nih.gov/research/NIHresearch/scunit/culture.asp

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Protocol for the culture of human embryonic stem cells (hESC). - [Read Culture of Human Embryonic Stem Cells (hESC) Protocol]

Histochemical Staining Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/H2.html

Category: Immunohistochemistry Protocols: Antibody Staining Protocols

Protocol for histochemical staining. This protocol has been optimized for b-galactosidase and human placental alkaline phosphatase staining in retinal tissue and cultured cells. - [Read Histochemical Staining Protocol]

Protocols for the Maintenance of Human Embryonic Stem Cells in Feeder Free Conditions - http://www.geron.com/PDF/scprotocols.pdf

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

Includes protocols: Mouse Embryonic Fibroblasts (MEF) Preparation; Harvesting MEFs; Cryopreservation of MEFs; Thawing and maintaining MEFs; Irradiating & Plating MEFs; Culture of Human ES cells with Matrigel® and Conditioned Medium; Preparation of Conditioned Medium (CM); Preparation of Matrigel® -coated plates; Passage of human ES cells on Matrigel®; Daily maintenance of feeder-free culture; Freezing Human ES Cells; Thawing Human ES cells; Formation of Embryoid Bodies; - [Read Protocols for the Maintenance of Human Embryonic Stem Cells in Feeder Free Conditions]

Human Embryonic Stem Cell Protocols - http://stemcells.nih.gov/staticresources/research/protocols/BresaGen_hESC_manual_2.1.pdf

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Human embryonic stem cells (hESCs) are pluripotent cells capable of differentiation to representatives of all three germ layers. Includes: Isolation of Primary Mouse Embryo Fibroblasts; Thawing and preparing p1 MEF feeder plates; Preparation of MEF- Conditioned Medium (MEF-CM; Microdissection Passaging of hESCs; Bulk passaging of hESC; Cryopreservation of hESCs; Thawing of hESCs; Karyotyping. - [Read Human Embryonic Stem Cell Protocols]

Radiation Safety Manual - http://www.cdc.gov/OD/ohs/manual/radman.htm

Category: Radioactivity: Radioaction Safety

Administration of the Radiation Safety Program, , Physical Security of Radioactive Materials, "As Low as Reasonably Achievable" (ALARA) Philosophy, Authorized Users, Radiation Workers , Permitted Workers, Training, Procurement, Inventory and Records, Personnel Monitoring, Radiation Exposure During Pregnancy, Laboratory Supplies and Equipment, Bioassay, Labeling, Surveys, Radioactive Waste Disposal. U.S. Department of Health and Human Services, CDC, Atlanta, GA. - [Read Radiation Safety Manual]

Purification of Human Mononuclear Cells and Neutrophils Protocol - http://www.cbrinstitute.org/labs/springer/protocols/chunduk_mono_neutro.html

Category: Immunology: Mononuclear Cell Isolation Protocols

Protocol for the purification of human mononuclear cells and neutrophils. - [Read Purification of Human Mononuclear Cells and Neutrophils Protocol]

Micronucleus assay Human Lymphocytes - http://ehs.sph.berkeley.edu/holland/protocolLibrary/Cytogenetics_MN_Lymphocytes.html

Category: Cytogenetics Protocols

MN in Human Lymphocytes (method and protocol description) Nina T. Holland, Division of Environmental Health Sciences, Berkeley - [Read Micronucleus assay Human Lymphocytes]

Fluorescence In Situ Hybridization of a Repetitive DNA Probe to Human Chromosomes in Suspension - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_104-107.pdf

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Protocol for fluorescence in situ hybridization of a repetitive DNA probe to human chromosomes in suspension. Hybridization technique which does not need formamide and dextran sulfate. As a model system, we used the repetitive specific human DNA probe pUC 1.77, labeled it with digoxigenin-11-dUTP by nick-translation, and hybridized it to metaphase chromosomes in suspension. These chromosomes were isolated by standard techniques from human lymphocytes. - [Read Fluorescence In Situ Hybridization of a Repetitive DNA Probe to Human Chromosomes in Suspension]

Culture of Human Prostatic Carcinoma Cell Lines - http://www.celldeath.de/apometh/rpmi.html

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]

A high-capacity RNA affinity column for the purification of human IRP1 and IRP2 overexpressed - http://www.rnajournal.org/cgi/reprint/9/3/364.pdf

Category: Chromatography Protocols: DNA RNA Affinity Chromatography

PDF. A high-capacity RNA affinity column for the purification of human IRP1 and IRP2 overexpressed in Pichia pastoris. CHARLES R. ALLERSON, ALAN MARTINEZ, EMINE YIKILMAZ and TRACEY A. ROUAULT. RNA Journal. - [Read A high-capacity RNA affinity column for the purification of human IRP1 and IRP2 overexpressed]

Articles (1-3 of 3)

Genomic DNA Labeling of Microarrays Protocol

DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.

Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA

This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.