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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]

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.]

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]

Applications of SKY in Cancer Cytogenetics Review - http://www.utoronto.ca/cancyto/protocols_software/cytogenetics/SKY.html

Category: Genetics and Genomics: Cancer Genetics Protocols

SKY has also been applied for the mouse genome, enabling investigators to extrapolate information from mouse models of cancer to their human counterparts. This review will address the advances that SKY has facilitated in the field of cancer cytogenetics, as well as its variety of application in the cancer research laboratories. - [Read Applications of SKY in Cancer Cytogenetics Review]

Calcium Dependent by Pentraxins to Ligands, and Effect of C-reactive Protein on Leptin Action - http://www.natureprotocols.com/2007/01/24/calcium_dependent_by_pentraxin.php

Category: Mouse Protocols

Protocol for calcium dependent by pentraxins to ligands immobilized on agarose, and effect of C-reactive protein on leptin action in mice in vivo. Includes: Testing for binding of CRP to immobilized leptin; Testing for binding of radiolabelled leptin by immobilized CRP, SAP and leptin receptor; Testing for effect of human CRP on human leptin action in mice. - [Read Calcium Dependent by Pentraxins to Ligands, and Effect of C-reactive Protein on Leptin Action]

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]

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]

Cell Culture Phototoxicity Test Protocol - http://embryo.ib.amwaw.edu.pl/invittox/prot/44.htm

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cytotoxicity Assays in Cell Culture Protocols

Human A431 cells and mouse 3T3 cells are exposed in culture to UV light both in the presence and absence of test compound. Phototoxicity is expressed as a decrease in cell viability as determined by the MTT assay. - [Read Cell Culture Phototoxicity Test Protocol]

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]

Contrast in Optical Microscopy - http://microscopy.fsu.edu/primer/techniques/contrast.html

Category: Microscopy Protocols: Optical Microscopy Protocols

When imaging specimens in the optical microscope, differences in intensity and/or color create image contrast, which allows individual features and details of the specimen to become visible. Contrast is defined as the difference in light intensity between the image and the adjacent background relative to the overall background intensity. In general, a minimum contrast value of 0.02 (2 percent) is needed by the human eye to distinguish differences between the image and its background. - [Read Contrast in Optical Microscopy]

Cryptic Translocation Identification in Human and Mouse Using Several Telomeric Multiplex FISH - http://info.med.yale.edu/genetics/ward/tavi/TMFISH.html

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Protocol for cryptic translocation identification in human and mouse using several telomeric multiplex FISH (TM-FISH) strategies. - [Read Cryptic Translocation Identification in Human and Mouse Using Several Telomeric Multiplex FISH]

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]

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)]

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]

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]

Deglycosylation of Glycoproteins Using Endoglycosidases - http://www.glycotech.com/protocols/Proto3.html

Category: Molecular Biology Protocols: Carbohydrate Protocols

Deglycosylation steps are different with every glycoprotein and must be determined empirically (Methods in Enzymology, 1994, 230: 44-57). A typical test reaction of human transferrin is described. T.H. Plummer, Jr. and A.L. Tarentino, Department of Biochemistry, New York State Department of Health, Wadsworth Center for Laboratories and Research, Albany, New York - [Read Deglycosylation of Glycoproteins Using Endoglycosidases]

Detection of Alu by PCR - http://www.accessexcellence.com/AE/AEPC/DNA/detection.html

Category: DNA Protocols: PCR Polymerase Chain Reaction Protocols

Detection of Alu by PCR - A Human DNA Fingerprinting Lab Protocol - 1994 Cold Spring Harbor Laboratory DNA Learning Center - [Read Detection of Alu by PCR]

Detection of Alu by PCR - http://www.accessexcellence.com/AE/AEPC/DNA/detection.html

Category: PCR Protocols

Detection of Alu by PCR - A Human DNA Fingerprinting Lab Protocol - 1994 Cold Spring Harbor Laboratory DNA Learning Center - [Read Detection of Alu by PCR]

Detection of mRNAs on Cryosections of the Cardiovascular System Using DIG-Labeled RNA Probes - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_189-196.pdf

Category: Cytogenetics Protocols: In Situ Hybridization Tissue Sections Protocols

Protocol for detection of mRNAs on cryosections of the cardiovascular system using DIG-labeled RNA probes. Protocol was optimized from a protocol using 35S-labeled RNA probes. It allows to detect the expression of low abundant mRNAs in the cardiovascular system, e.g. of the proinflammatory cytokine GM-CSF in normal human coronary arteries, and of IL6 and gp130 in human failing hearts. The protocol can be combined with immunohistochemistry. - [Read Detection of mRNAs on Cryosections of the Cardiovascular System Using DIG-Labeled RNA Probes]

Determination of Neopterin and Biopterin by Liquid Chromatography Protocol - http://www.natureprotocols.com/2006/10/13/determination_of_neopterin_and.php

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocol for determination of neopterin and biopterin by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) in rat and human plasma, cell extracts and tissue homogenates. - [Read Determination of Neopterin and Biopterin by Liquid Chromatography Protocol]

Direct Selection of cDNAs with Large Genomic DNA Clones Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4077

Category: Genetics and Genomics: Genotyping Protocols: Mutation Detection Protocols

The goal of this method is to identify transcriptionally active genes in cloned segments of genomic DNA. The protocol uses hybridization and affinity purification to recover biotin-labeled cDNAs that bind to a 500-kb segment of human DNA cloned in a BAC vector. However, the method can be easily adapted to other clones of genomic DNAs cloned in high-capacity vectors. - [Read Direct Selection of cDNAs with Large Genomic DNA Clones Protocol]

Direct Sequencing Using MultiPlex PCR-Based Method - http://www.genome.ou.edu/MultiplexPCRbasedSeq.html

Category: DNA Protocols: DNA Sequencing Protocols

Direct sequencing from amplified bacterial and large insert cloned human or mouse genomic DNA via an improved MultiPlex PCR-based method. Includes: Preparing primers for MP-PCR; Amplification; PCR Product Clean-up; Sequencing. - [Read Direct Sequencing Using MultiPlex PCR-Based Method]

DNA In Situ Hybridization with an Alkaline Phosphatase-Based Fluorescent Detection System Protocol - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_124-126.pdf

Category: Nucleic Acid Modification Protocols: DIG Biotin Labeled Probes Protocols

Protocol describes a high sensitivity indirect detection procedure for DIG-labeled hybridization probes. The procedure uses the components of the HNPP Fluorescent Detection Set to form a fluorescent precipitate of HNPP (2-hydroxy-3-naphthoic acid-2’-phenylanilide phosphate) and Fast Red TR at the site of hybridization. This procedure can be used to detect single copy sequences as small as 1 kb on human metaphase chromosomes. - [Read DNA In Situ Hybridization with an Alkaline Phosphatase-Based Fluorescent Detection System 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]

Feeder Cell Density - A key Parameter in Human Embryonic Stem Cell Culture - http://www.bioone.org/bioone/?request=get-document&issn=1543-706X&volume=040&issue=08&page=0255

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

Feeder Cell Density - A key Parameter in Human Embryonic Stem Cell Culture - [Read Feeder Cell Density - A key Parameter in Human Embryonic Stem Cell Culture]

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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.