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CaPO4 Transfection for Chromaffin Cells - http://medicine.ucsd.edu/hypertension/list_of_protocols/15_CaPO4_Transfect_sw.htm

Category: Cell Biology and Cell Culture: Cell Transfection Protocol

Transfection of DNA into cells using PIBS, DNA, CaCl2 and Calcium phosphate. Daniel O'Connor, UCSD Chromaffin Cell and Hypertension Research - [Read CaPO4 Transfection for Chromaffin Cells]

Carbohydrate-Specific Adhesion of Intact Cells to Resolved Glycolipids on TLC Plates - http://www.glycotech.com/protocols/Proto9.html

Category: Molecular Biology Protocols: Carbohydrate Protocols

Protocol detects specific cell adhesion to glycolipids resolved on TLC plates. Carbohydrate-Specific Adhesion of Intact Cells to Resolved Glycolipids on TLC Plates. Ronald L. Schnaar~Professor, Johns Hopkins University Medical School, Baltimore, Maryland. Glycotech. - [Read Carbohydrate-Specific Adhesion of Intact Cells to Resolved Glycolipids on TLC Plates]

cDNA Synthesis and Real-Time PCR Using RNA from Laser-Captured Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4108

Category: PCR Protocols: Real-Time PCR Protocols

Protocol describes the real-time PCR using RNA purified from laser-captured tissues Laser Capture Microdissection (LCM): Preparation and Sectioning of Frozen Tissue Blocks and Purification of RNA from Isolated Cells. - [Read cDNA Synthesis and Real-Time PCR Using RNA from Laser-Captured Cells Protocol]

cDNA Synthesis from MOLT-4 Cells - http://www.musc.edu/BCMB/ceramide/protocols/0043.html

Category: RNA Protocols: RT-PCR and cDNA synthesis

cDNA Synthesis from MOLT-4 Cells. First strand of cDNA synthsis. Cungui Mao - [Read cDNA Synthesis from MOLT-4 Cells]

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]

Cell Fractionation of a Mixed Population of Cells - http://www.axis-shield.com/densityhome/optiprep/C14.pdf

Category: Cell Biology and Cell Culture

Cell fractionation techniques are presented for the design of gradient systems for separating one or more cell types from lavages of body cavities or from mechanically or enzymically-dissociated tissues. Includes: Preparation of cell suspension for gradient loading; Fractionation by buoyant density; Fractionation on the basis of cell size. - [Read Cell Fractionation of a Mixed Population of Cells]

Cell Fractionation protocol Usingt Low Speed Separaton of Cells - http://homepages.gac.edu/~cellab/chpts/chpt3/ex3-4.html

Category: Cell Biology and Cell Culture

Cell fractionation protocol using low speed seperation. - [Read Cell Fractionation protocol Usingt Low Speed Separaton of Cells]

Cell Freezing using Liquid Nitrogen N2 - http://www.liv.ac.uk/~giles/methods/cell_freezing.htm

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

Freezing gealthy cells (> 95% viable) the protocol obtains a culture 80-90% viable 24 hours after thawing and growing on test vial from step 9. Antisense Research Group - [Read Cell Freezing using Liquid Nitrogen N2]

Cell Staining for Immunofluorescence Microscopy - http://www.bdbiosciences.com/pharmingen/protocols/Immunofluorescence_Microscopy.shtml

Category: Molecular Imaging: Immunofluorescence Microscopy

Cell Staining for Immunofluorescence Microscopy. Includes protocols for fixing the cells, Coverslip Preparation for Adherent Cells, Coverslip Preparation for Non-Adherent Cells, Paraformaldehyde Fixation, and Methanol/Acetone Fixation. Blocking protocols include blocking with primary antibody, and incubation with secondary antibody. - [Read Cell Staining for Immunofluorescence Microscopy]

Cell Staining for Sorting of Hematopoietic Stem Cells (HSC) and Myeloid Progenitors - http://www.natureprotocols.com/2006/06/29/cell_staining_for_sorting_of_h.php

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Sorting Protocols

The combination of prospective identification/isolation of bone marrow progenitors and quantitative RT-PCR is a powerful tool to understand the molecular mechanism underlying hematopoiesis. Describes the standard procedures of the murine myeloid progenitor staining for fluorescence activated cells sorting (FACS) and RNA purification methods. - [Read Cell Staining for Sorting of Hematopoietic Stem Cells (HSC) and Myeloid Progenitors]

Cell Staining for Sorting of Hematopoietic Stem Cells and Myeloid Progenitors and Isolating RNA - http://www.natureprotocols.com/2006/06/29/cell_staining_for_sorting_of_h.php

Category: Stem Cell Protocols

The combination of prospective identification/isolation of bone marrow progenitors and quantitative RT-PCR is a powerful tool to understand the molecular mechanism underlying hematopoiesis. Here, we described our standard procedures of the murine myeloid progenitor staining for fluorescence activated cells sorting (FACS) and RNA purification methods. - [Read Cell Staining for Sorting of Hematopoietic Stem Cells and Myeloid Progenitors and Isolating RNA]

Cell Synchronization Using Centrifugal Elutriation Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4490

Category: Cell Biology and Cell Culture: Cell Cycle Protocols

protocol describes a method for the synchronization of cell populations using centrifugal elutriation. The method relies on the fact that cell size increases linearly as cells proceed through the cell cycle. Cells of similar size (and cell cycle phase) are eluted stepwise from the cell chamber, with the smallest size (those in early G1) being eluted first. Using this procedure, it is possible to obtain relatively pure populations of cells at various points in G1, S, and G2/M. - [Read Cell Synchronization Using Centrifugal Elutriation Protocol]

Cell Synchronization Using Centrifugal Elutriation Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4490

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Cycle Analysis Protocols

This protocol describes a method for the synchronization of cell populations using centrifugal elutriation. The method relies on the fact that cell size increases linearly as cells proceed through the cell cycle. Cells of similar size (and cell cycle phase) are eluted stepwise from the cell chamber, with the smallest size (those in early G1) being eluted first. Using this procedure, it is possible to obtain relatively pure populations of cells at various points in G1, S, and G2/M. - [Read Cell Synchronization Using Centrifugal Elutriation Protocol]

Cell Thawing and cell line Freezing Protocol - http://iacf.bsd.uchicago.edu/Monoclonal_Antibody_Facility/FreezeThawProtocol.htm

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

Method for freezing and thawing cells. Immunology Applications Core Facility. U. Chicago - [Read Cell Thawing and cell line Freezing Protocol]

Cell Viability Assays that Measure ATP Protocol - http://www.promega.com/paguide/chap4.htm#title2

Category: Cell Biology and Cell Culture: Cell Viability Protocols

The CellTiter-Glo® Luminescent Cell Viability Assay is a homogeneous method to determine the number of viable cells in culture. Detection is based on using the luciferase reaction to measure the amount of ATP from viable cells. The amount of ATP in cells correlates with cell viability. - [Read Cell Viability Assays that Measure ATP Protocol]

Cell Viability Assays that Measure Metabolic Capacity Protocol - http://www.promega.com/paguide/chap4.htm#title3

Category: Cell Biology and Cell Culture: Cell Viability Protocols

The CellTiter-Blue® Cell Viability Assay uses an optimized reagent containing resazurin. The homogeneous procedure involves adding the reagent directly to cells in culture at a recommended ratio of 20µl of reagent to 100µl of culture medium. - [Read Cell Viability Assays that Measure Metabolic Capacity Protocol]

CellTiter-Glo Luminescent Cell Viability Assay Protocol - http://www.promega.com/tbs/tb288/tb288.pdf

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

Protocol for CellTiter-Glo luminescent cell viability assay. This assay is a homogeneous method of determining the number of viable cells in culture based on quantitation of the ATP present, which signals the presence of metabolically active cells. - [Read CellTiter-Glo Luminescent Cell Viability Assay Protocol]

Chambers for Examination of Live Cells under Mechanical Stress Protocol - http://www.cshprotocols.org/cgi/content/extract/2007/3/pdb.ip32

Category: Microscopy Protocols: Live-Cell Imaging Protocols

Manual measurement and manipulation of the cell surface requires access to the cells, usually in an open chamber. Temperature-controlled chambers or stage inserts are preferred for maintaining physiological activity during the experiment. For example, heated culture dishes with coverslip glass bottoms (Bioptechs) permit high-resolution fluorescence microscopy of living cells during force application. - [Read Chambers for Examination of Live Cells under Mechanical Stress Protocol]

Characterization of Cells by Flow Cytometry Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000018.pdf?pid=PP00000018

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol uses specific antibodies coupled to one of four fluorochromes: fluorescein isothiocyanate (FITC), R-phycoerythrin (PE), peridinin chlorophyll-a (PcP), and allophycocyanin (APC). These fluorochromes can be used simultaneously to stain and analyze the expression patterns of four different proteins in the same sample. The fluorochrome stained cell populations are analyzed using a FACSCalibur dual-laser flow cytometer. - [Read Characterization of Cells by Flow Cytometry Protocol]

Characterization of Cells by Flow Cytometry Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000018.pdf?pid=PP00000018

Category: Immunology: B Cell Protocols

Fluorochromes can be used simultaneously to stain and analyze the expression patterns of four different proteins in the same sample. The fluorochrome stained cell populations are analyzed using a FACSCalibur dual-laser flow cytometer. - [Read Characterization of Cells by Flow Cytometry Protocol]

Chemotaxis Assay - http://www.cbrinstitute.org/labs/springer/protocols/melissa_chemotaxis.html

Category: Immunology: Chemotaxis

Chemotaxis Assay, Springer Lab. A chemotaxis assay's function is to assess whether a factor or molecule of interest has chemotactic activity on a motile cell type. Chemotaxis is the ability of a factor to cause the migration of a cell. The chemotactic assay is based on the creation of a chemical gradient of the chemotactic agent which will cause cells to migrate through the gradient towards the chemotactic agent. - [Read Chemotaxis Assay]

Chemotaxis Assay Protocol - http://www.cbrinstitute.org/labs/springer/protocols/melissa_chemotaxis.html

Category: Cell Biology and Cell Culture

This chemotaxis assay protocol is based on the premise of creating a gradient of the chemotactic agent and allowing cells to migrate through a membrane towards the chemotactic agent. A chemotaxis assay can determine whether your protein or small molecule of interest has chemotactic activity on a specific cell type. Chemotaxis is then the ability of a protein to direct the migration of a specific cell. - [Read Chemotaxis Assay Protocol]

Chemotaxis Assay Using Microchemotaxis Chambers (Modified Boyden Chamber) - http://bioprotocol.bio.com/protocolstools/protocol.jhtml?id=p2158

Category: Signalling Signal Transduction Protocols: Chemokine Protocols

Simple apparatus to study chemotaxis of leukocytes or other migratory cells. Protocol for Chemotaxis. BioProtocol - [Read Chemotaxis Assay Using Microchemotaxis Chambers (Modified Boyden Chamber)]

ChIP Assay Protocol PDF - http://www.bcm.edu/rosenlab/protocols/ChIP.pdf

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

Formaldehyde cross-linking and chromatin immunoprecipitation assays of tissue culture cells, Based on Boyd and Farnham. Michelle Kallesen, Rosen Lab. - [Read ChIP Assay Protocol PDF]

ChIP ping for Dummies - http://mcardle.oncology.wisc.edu/sugden/Protocols/html%20files/ChIPping%20for%20Dummies.htm

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

Very nice and detailed ChIP assay protocol procedure. Blocking Staph A cells, Crosslinking cells, Chromatin preparation from cell pellets, sonication, blocking. Sugden Laboratory - [Read ChIP ping for Dummies]

Articles (1-10 of 24)

Lambda Phage Protocol Large DNA Preparation

Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

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.

Drosophila Cell Transfection Protocol

A simple Drosophila tissue culture cell transfection method.

Tubulin Polymerization Protocol using GTP and GMPCPP

Tubulin is polymerized into microtubules by incubating tubulin at 37°C with GTP. A nucleation seed is added when the purpose is to assay microtubule elongation. Tubulin can also be polymerized for the purposes of recycling the tubulin or labeling the microtubules with fluorescently labeled tubulin. Based on the protocol by Timothy Mitchison of Harvard University.

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.

Acid Guanidinium Thiocyanate RNA Isolation Phenol Chloroform Extraction

A single step RNA isolation protocol using Phenol Chloroform Extraction and Acid Guanidinium Thiocyanate. This RNA isolation method uses the fact that guanidinium thiocyanate can simultaneously lyse the cells and inactive cellular RNAses during the initial RNA isolation step allow a single step in the method.

Microinjection Pipettes Protocol

Microinjection Pipettes Preparation Protocol.

Acid Washing Microinjection Pipettes Protocol

Acid Washing of Glass microinjection pipettes protocol.

Paraffin Embedding Protocol

Paraffin Embedding Protocol for molecular profiling. This Paraffin Embedding Protocol describes the processing of the tissues into sections following ethanol fixation. Molecular profiling (MP) is a technique that is used to visualize the global patterns of RNA expression or protein expression in various cell types and disease processes.

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