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Chromatography, Gel: Chromatography on non-ionic gels without regard to the mechanism of solute discrimination.

Column Chromatography

JoVE 10217

Source: Laboratory of Dr. Jimmy Franco - Merrimack College

Column chromatography is one of the most useful techniques for purifying compounds. This technique utilizes a stationary phase, which is packed in a column, and a mobile phase that passes through the column. This technique exploits the differences in polarity between compounds, allowing the molecules to be facilely separated.1 The two most common stationary phases for column chromatography are silica gel (SiO2) and alumina (Al2O3), with the most commonly used mobile phases being organic solvents.2 The solvent(s) chosen for the mobile phase are dependent on the polarity of the molecules being purified. Typically more polar compounds require more polar solvents in order to facilitate the passage of the molecules through the stationary phase. Once the purification process has been completed the solvent can be removed from the collected fractions using a rotary evaporator to yield the isolated material.


 Essentials of Organic Chemistry

Isolation and Characterization Of Chimeric Human Fc-expressing Proteins Using Protein A Membrane Adsorbers And A Streamlined Workflow

1Department of Chemical and Biomolecular Engineering, Ohio University, 2Biomedical Engineering Program, Russ College of Engineering and Technology, Ohio University, 3Department of Dermatology, Brigham and Women's Hospital, Harvard Medical School

JoVE 51023


 Bioengineering

CN-GELFrEE - Clear Native Gel-eluted Liquid Fraction Entrapment Electrophoresis

1Departments of Chemistry and Molecular Biosciences, Chemistry of Life Processes Institute, Proteomics Center of Excellence, Robert H. Lurie Comprehensive Cancer Center, Northwestern University, 2Institute of Chemistry, Proteomics Unit, Federal University of Rio de Janeiro, 3Department of Cell Biology, Brazilian Center for Protein Research, Laboratory of Biochemistry and Protein Chemistry, University of Brasilia

JoVE 53597


 Chemistry

Purification of a Total Lipid Extract with Column Chromatography

JoVE 10159

Source: Laboratory of Jeff Salacup - University of Massachusetts Amherst

The product of an organic solvent extraction, a total lipid extract (TLE), is often a complex mixture of hundreds, if not thousands, of different compounds. The researcher is often only interested in a handful of compounds. The compounds of interest may belong to one of several classes of compounds, such as alkanes, ketones, alcohols, or acids (Figure 1), and it may be useful to remove the compound classes to which it does not belong in order to get a clearer view of the compounds you are interested in. For example, a TLE may contain 1,000 compounds, but the Uk'37 sea surface temperature proxy is based on only two compounds (alkenones) and the TEX86 sea surface temperature proxy is based on only four (glycerol dialkyl glycerol tetraethers). It would behoove the researcher to remove as many of the compounds they are not interested in. This makes the instrumental analysis of the compounds of interest (alkenones or GDGTs) less likely to be complicated by other extraneous compounds. In other cases, an upstream purification technique may have produced compounds you wish to now remove from the sample, such as the production of carboxylic acids during saponification in our


 Essentials of Earth Science

High Resolution Quantitative Synaptic Proteome Profiling of Mouse Brain Regions After Auditory Discrimination Learning

1Leibniz Institute for Neurobiology (LIN), 2Institute of Experimental Internal Medicine, Medical School, Otto von Guericke University Magdeburg, 3Institute of Pharmacology and Toxicology, Medical School, Otto von Guericke University

JoVE 54992


 Neuroscience

Modified Terminal Restriction Fragment Analysis for Quantifying Telomere Length Using In-gel Hybridization

1Departments of Pathology and Infectious Diseases and Microbiology, University of Pittsburgh, 2University of Pittsburgh Cancer Institute, 3Department of Environmental and Occupational Health, University of Pittsburgh, 4Departments of Psychiatry, Psychology, Behavioral & Community Health Sciences, University of Pittsburgh

JoVE 56001


 Cancer Research

From Constructs to Crystals – Towards Structure Determination of β-barrel Outer Membrane Proteins

1Department of Biological Sciences, Markey Center for Structural Biology, Purdue University, 2National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of Health, 3National Institute of General Medical Sciences (NIGMS), National Institutes of Health

JoVE 53245


 Chemistry

Optimized Protocol for the Extraction of Proteins from the Human Mitral Valve

1Centro Cardiologico Monzino IRCCS, 2Cardiovascular Tissue Bank of Milan, Centro Cardiologico Monzino IRCCS, 3Department of Clinical Sciences and Community Health, Cardiovascular Section, University of Milan, 4Department of Cardiovascular Disease, Development and Innovation Cardiac Surgery Unit, Centro Cardiologico Monzino IRCCS

JoVE 55762


 Biochemistry

Initial Evaluation of Antibody-conjugates Modified with Viral-derived Peptides for Increasing Cellular Accumulation and Improving Tumor Targeting

1Department of Nuclear Medicine and Radiobiology, Université de Sherbrooke, 2Sherbrooke Molecular Imaging Center (CIMS), Université de Sherbrooke, 3Sherbrooke Institute of Pharmacology

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JoVE 55440


 JoVE In-Press

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