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A Method for Micro-Scale Isolation and Purification of Gangliosides - http://www.glycotech.com/protocols/Proto1.html

Category: Molecular Biology Protocols: Carbohydrate Protocols

Solvent partition protocol allows the isolation of gangliosides from small samples and from samples where ganglioside concentrations are low, especially relative to the concentration of potentially contaminating proteins and other large molecular weight species. Stephan Ladisch Director, Center for Cancer and Transplant Biology, Children's National Medical Center, Washington, D.C. - [Read A Method for Micro-Scale Isolation and Purification of Gangliosides]

Absorbance Assay 280 nm - http://www.ruf.rice.edu/~bioslabs/methods/protein/abs280.html

Category: Protein Protocols: Protein Quantitation Concentration Protocols

Absorbance assays are fast and convenient, since no additional reagents or incubations are required. No protein standard need be prepared. The assay does not consume the protein. The relationship of absorbance to protein concentration is linear. Because different proteins and nucleic acids have widely varying absorption characteristics there may be considerable error, especially for unknowns or protein mixtures. - [Read Absorbance Assay 280 nm]

Annexin V Protocol for Flow Cytometry - http://www.niehs.nih.gov/research/atniehs/core/fc/protocols/annexin.cfm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Different cell types vary in their phosphatidylserine (PS) content, along with the amount of PS exposure on the cell surface after cell death. This protocol is a guideline for getting started, however it may be necessary to adjust the concentration of the Annexin V-FITC. - [Read Annexin V Protocol for Flow Cytometry]

Antibiotic Concentration in Media - http://wheat.pw.usda.gov/~lazo/methods/lazo/antibiot.html

Category: Buffers Media, and Solutions: Bacterial Solutions

Antibiotic Concentration in Media. (Gerard R. Lazo) - [Read Antibiotic Concentration in Media]

Bradford Assay Method - http://research.cm.utexas.edu/bkitto/Kittolabpage/Protocols/Biochemistry/Bradford.html

Category: Protein Protocols: Protein Quantitation Concentration Protocols: Bradford Protein Assay Protocols

The bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie Brilliant blue to protein. There is no interference from cations nor from carbohydrates such as sucrose.
Detergents such as sodium dodecyl sulfate and triton x-100 can interfere with the assay, as well as strongly alkaline solutions.

Includes a general overview of the procedure and preparation of the standards in the protocol. - [Read Bradford Assay Method]

Bradford Protein Assay Spectrophotometry - http://homepages.gac.edu/~cellab/appds/appd-g.html#exG-1

Category: Protein Protocols: Protein Quantitation Concentration Protocols: Bradford Protein Assay Protocols

Bradford Protein Assay Spectrophotometry. Includes spectrophotometry information and the Bradford protein assay: A spectrophotometer or colorimeter makes use of the transmission of light through a solution to determine the concentration of a solute within the solution. A spectrophtometer differs from a colorimeter in the manner in which light is separated into its component wavelengths. A spectrophotometer uses a prism to separate light and a colorimeter uses filters. - [Read Bradford Protein Assay Spectrophotometry]

Bradford Protein Concentration Assay - http://web.chemistry.gatech.edu/~williams/bCourse_Information/4581/techniques/bradford/bradford.html

Category: Protein Protocols: Protein Quantitation Concentration Protocols: Bradford Protein Assay Protocols

Includes Abbreviations, Background, and Procedure steps using BSA. The Bradford protein assay (1) is one of several simple methods commonly used to determine the total protein concentration of a sample. The method is based on the proportional binding of the dye Coomassie to proteins. The assay is colorimetric; as the protein concentration increases, the color of the test sample becomes darker. Coomassie absorbs at 595 nm. - [Read Bradford Protein Concentration Assay]

Cajal Body Isolation Protocol - http://www.lamondlab.com/f7cbprotocol.htm

Category: Cell Organelle Protocols: Nucleus Protocols

Cajal Body Isolation Protocol. Protocol includes: Sonication, Removal Nucleoi, Gradient One, Gradient two, Concentration and final enrichment of cajal bodies. Also includes: Making 2.55M sucrose stock and Analysis of the enriched Cajal body fraction. - [Read Cajal Body Isolation Protocol]

Concentrating and Desalting Nucleic Acids with Microconcentrators Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4457

Category: Nucleic Acid Purification Protocols

Ultrafiltration is an alternative to ethanol precipitation for the concentration and desalting of nucleic acid solutions. It requires no phase change and is particularly useful for dealing with very low concentrations of nucleic acids. This protocol describes the use of the Microcon cartridge, a centrifugal ultrafiltration device, to concentrate and desalt nucleic acid solutions. - [Read Concentrating and Desalting Nucleic Acids with Microconcentrators Protocol]

Concentration of Oligo DNA by Ethanol Precipitation Protocol - http://mycoplasmas.vm.iastate.edu/lab_site/methods/DNA/precip.html

Category: Oligonucleotide Protocols: Oligonucleotide Purification Protocols

Concentration of DNA by Ethanol Precipitation Protocol. Adapted from Bruce A. Roe, Department of Chemistry and Biochemistry, The University of Oklahoma, Norman, Oklahoma. Usually 2.5 - 3 volumes of ethanol and/or acetate solution is added to the DNA in a microcentrifuge tube. This is then put into an ice-water bath for at least 10 minutes. The precipitation is performed by incubation at -20C overnight. - [Read Concentration of Oligo DNA by Ethanol Precipitation Protocol]

Cytokine ELISA Protocol - http://www.bdbiosciences.com/pharmingen/protocols/Cytokine_ELISA.shtml

Category: ELISA Protocols: Sandwich ELISA Assay Protocols

Cytokine sandwich ELISA are sensitive enzyme immunoassays that can specifically detect and quantitate the concentration of soluble cytokine and chemokine proteins. BD Biosciences - [Read Cytokine ELISA Protocol]

Determination of the Extinction Coefficient for a Protein of Unknown Concentration - http://www.ruf.rice.edu/~bioslabs/methods/protein/absext.html

Category: Protein Protocols: Protein Quantitation Concentration Protocols

Determination of the Extinction Coefficient for a Protein of Unknown Concentration. The concentration can be determined for a solution of a pure protein with unknown extinction coefficient. - [Read Determination of the Extinction Coefficient for a Protein of Unknown Concentration]

DNA IQ Isolation of Genomic DNA from Stains and Buccal Swabs Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4098

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: Genomic DNA Isolation Protocols

In this protocol, the DNA-binding capacity of Wizard MagneSil particles is used to capture and release a consistent amount of DNA (100 ng) across a wide range of samples. At the end of the procedure, the DNA is eluted into 100 µl Elution Buffer to give a final concentration of 1 ng/µl, relieving the need for postpurification DNA quantitation. - [Read DNA IQ Isolation of Genomic DNA from Stains and Buccal Swabs Protocol]

DNA-Methyltransferase Assay Protocol - http://www.mdanderson.org/departments/methylation/display.cfm?id=9B5B6082-C53A-4A86-BEE3EE149A52251E&method=displayFull&pn=A3F15D82-C9B4-41CD-BA4BE767E50A73D2

Category: Nucleic Acid Modification Protocols: DNA Methylation Protocols

Protocol for DNA-methyltransferase assay. Includes: Homogenize Cells/Tissues; Quantitate Protein Concentration; Dilute Samples; The Assay Proper; Troubleshooting. - [Read DNA-Methyltransferase Assay Protocol]

Dye Protocols and Notes for Cosmid, BAC, BAC, Fosmid Templates - http://www.genome.ou.edu/big_dyes_cos_bac_pac_fos.html

Category: BACs Bacterial Artificial Chromosome Protocols

Sequencing conditions tested for the ABI Big-Dye terminators (PE-ABI #4303150 for the 1000 reaction kit - Description: TF,KIT BTD RR-1000) with various templates. Important to quantitate all templates by agarose gel electrophoresis vs size and concentration standards and do a few tests with different template concentrations to determine the optimal conditions for your reactions. Several conditions are given. - [Read Dye Protocols and Notes for Cosmid, BAC, BAC, Fosmid Templates]

Early Events in B Lymphocyte Activation Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E6634190D367003ECF94AE65295FA77&objectid=6674AB8295693316922D8DFC029193A6

Category: Immunology: B Cell Protocols

B cell activation can be quantitated indirectly by assaying antibody production or directly by measuring cellular changes that occur immediately after exposure to an activation signal. Provides methods for the latter (direct) approach--namely, methods for quantifying early parameters of B cell activation such as increases in intracellular ionized calcium concentration [Ca2+]I, cell size, and MHC class II-antigen expression. - [Read Early Events in B Lymphocyte Activation Protocol]

Extraction and Purification of Total RNA using TRIzol OR TRI Reagent Protocol - http://www.animal.ufl.edu/hansen/Protocols/RNA_extraction.htm

Category: Nucleic Acid Purification Protocols: RNA Isolation Protocols

Protocol for extraction and purification of total RNA using TRIzol OR TRI reagent. Includes: Homogenization for Cell Suspensions; Phase Separation; RNA Precipitation; RNA Wash; Redissolving the RNA; Determination of RNA Concentration and Purity; Preparation of Rnase-free water. - [Read Extraction and Purification of Total RNA using TRIzol OR TRI Reagent Protocol]

Fluorometric Quantitation of DNA Using Hoechst 33258 Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4458

Category: Nucleic Acid Detection Protocols

Protocol describes the quantitation of DNA using Hoechst 33258, a fluorescent dye that binds to double-stranded DNA. Fluorometry is simple and more sensitive than spectrophotometry, and allows the detection of nanogram quantities of DNA. The assay can only be used to measure the concentration of DNAs whose sizes exceed ~1 kb, as Hoechst 33258 binds poorly to smaller DNA fragments. - [Read Fluorometric Quantitation of DNA Using Hoechst 33258 Protocol]

Growth and Concentration of Chlorogonium for Culturing Freshwater Hypotrichs Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4464

Category: Laboratory Model Organisms Protocols

Protocol describes the growth and concentration of the alga Chlorogonium elongatum as a food source for culturing freshwater hypotrichs. Most freshwater hypotrichs (including Oxytricha nova, O. fallax, and O. trifallax; Euplotes aediculatus and E. eurystomous; and Stylonychia lemnae) can be grown to high density with Chlorogonium as the food organism. A similar regimen can be used to prepare other food sources such as Tetrahymena or bacteria (e.g., Aerobacter aerogenes). - [Read Growth and Concentration of Chlorogonium for Culturing Freshwater Hypotrichs Protocol]

Growth and Purification of 25-100 ug Lambda Clone DNA Protocol - http://humgen.wustl.edu/hdk_lab_manual/lambda/lambda13.html

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

There are essentially three parts to this protocol: 1. growth of at least 5x10e8 pfu phage to provide an inoculum growth of a larger liquid lysate that will produce about 5x10e12 pfu; 2. concentration and purification of the phage, and; 3. DNA preparation. - [Read Growth and Purification of 25-100 ug Lambda Clone DNA Protocol]

GUS Reporter Gene Assay Protocol - http://www.oardc.ohio-state.edu/stockingerlab/Protocols/GUS%20Assays.pdf

Category: Reporter Gene Assays

Protocol for GUS reporter gene assay. Includes: Protein isolation; Alternative method for small (<1g) quantities of tissue; GUS assays; Bradford protein concentration determination assays - [Read GUS Reporter Gene Assay Protocol]

In Vitro Endoplasmic Reticulum to Golgi Transport Reaction in Yeast Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p1831

Category: Yeast Protocols

Protocol for in vitro endoplasmic reticulum to golgi transport reaction in Yeast. Includes: Preparation of Membranes; One-Stage Reaction; Two-Stage Reaction using Normal Amount of Membranes; Two-Stage Reaction using Low Concentration of Membranes. - [Read In Vitro Endoplasmic Reticulum to Golgi Transport Reaction in Yeast Protocol]

Inducing Yeast Cell Synchrony: alpha-Factor Arrest Using Low Cell Concentration Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4171

Category: Yeast Protocols: Yeast Cell Synchrony Protocols

It is sometimes desirable to have an entire population of yeast cells that is growing synchronously or is arrested at a unique point in the cell cycle. - [Read Inducing Yeast Cell Synchrony: alpha-Factor Arrest Using Low Cell Concentration Protocol]

Kit for Rapid Enrichment of SARS Virus - http://www.gentaur.com/sars_isolation.htm

Category: Viral Culture and Isolation Protocols: Viral Isolation Protocols

During incubation period and the course of disease, SARS virus replicates and releases from cells. The level of virus varies greatly from sample to sample. At early stage of incubation period or during the late convalescent phase, the concentration of virus is very low in all kinds of samples. Virus is also detected at extremely low concentrations in plasma during... - [Read Kit for Rapid Enrichment of SARS Virus]

List of Protein Assays - http://www.ruf.rice.edu/~bioslabs/methods/protein/protein.html

Category: Protein Protocols: Protein Quantitation Concentration Protocols

Find a list of assays for the determination of protein concentration in a solution. This list includes the sensitivity range, volume/amount of sample needed, subjective comments on accuracy and convenience, and major interfering agents. Procedural details, equipment requirements, and references are outlined in the individual assay documents. - [Read List of Protein Assays]

Articles (1-7 of 7)

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.

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.

In Vitro Translated Xenopus Mos Kinase Assay Protocol

In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.

Coupling Cysteine Containing Peptides to Carrier Protein for Immunization and Polyclonal Antibody

This protocol is used in the production of polyclonal antibodies that recognize a peptide sequence of interest.

Absorption Spectroscopy and Quantification of Filamentous Phage Protocol

Unlike spherical phage, such as T4 and λ, which have roughly equal weight ratios of protein to DNA, filamentous phage have about six times more protein than DNA; the protein therefore contributes substantially to the absorption spectrum.

Picking Transfected ES Cell Clones Protocol

This protocol a protocol on how to generate transfected embryonic stem (ES) cell clones. The previous protocol in this series is the Protocol for Electroporation of ES cells. The next protocol in the series is the Protocol on Disaggregation, Expansion, and Freezing of Transfected ES Clones.

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