The Journal of Visualized Experiments (JoVE) is a peer reviewed, PubMed-indexed video journal. Our mission is to increase the productivity of scientific research.

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Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification


JoVE 3060 9/21/2011

1College of Nursing, Interdisciplinary Life Sciences Research Laboratory, Seattle University, 2College of Science and Engineering, Interdisciplinary Life Sciences Research Laboratory, Seattle University

An automated method for identifying suitable hydrophobic interaction chromatography (HIC) media to be used in the process of protein purification is presented. The method utilizes a medium-pressure liquid chromatography system including automated buffer blending, dynamic sample loop injection, sequential column selection, multi-wavelength analysis, and split fraction eluate collection.

 

Basic Principles for Purification Using Supercritical Fluid Chromatography - ADVERTISEMENT


JoVE 2604 11/11/2010

Chemistry Commercial Operations, Waters Corporation

Basic principles for the separation of compounds from mixtures using supercritical fluid chromatography (SFC) are similar to the fundamentals of preparative liquid chromatography.

 

Analytical HPLC to Preparative HPLC: Scale Up Techniques using a Natural Product Extract - ADVERTISEMENT


JoVE 2885 8/11/2011

Pharmaceutical Business Operations, Waters Corporation

Using the Waters AutoPurification™ System, separation methods can be developed on an analytical scale and transferred to preparatory scale on the same system.

 

Use of Arabidopsis eceriferum Mutants to Explore Plant Cuticle Biosynthesis


JoVE 709 5/31/2008

1Department of Botany, University of British Columbia - UBC, 2Department of Chemistry, University of British Columbia - UBC

The plant cuticle is a waxy outer covering on plants that has a primary role in water conservation but is also an important barrier against the entry of pathogenic microorganisms. In this video, we demonstrate the analysis of plant cuticle mutants identified by forward and reverse genetics approaches.

 

A Protocol for the Production of KLRG1 Tetramer


JoVE 1701 1/12/2010

Department of Molecular Microbiology and Immunology, Brown University

This protocol describes the production of KLRG1 tetramer, which is a powerful tool for the analysis of KLRG1 ligands.

 

Purification of the M. magneticum Strain AMB-1 Magnetosome Associated Protein MamAΔ41


JoVE 1844 3/25/2010

Department of Life Sciences and National Institute for Biotechnology in the Negev, Ben-Gurion University

MamA is a unique Magnetosome associated protein which was shown to be involved in magnetosome activation. Here we present the purification protocol of MamA deletion mutant (MamAΔ41) from M. magneticum AMB-1.

 

A Lectin HPLC Method to Enrich Selectively-glycosylated Peptides from Complex Biological Samples


JoVE 1398 10/01/2009

1Obstetrics, Gynecology and Reproductive Sciences, University of California, San Francisco - UCSF, 2Buck Institute for Age Research, 3Department of Chemistry, Purdue University

Lectin-conjugated POROS beads were employed for HPLC. Glycopeptide standards served as positive and negative controls. MARS-14 depleted, trypsin-digested human plasma was chromatographed and flow-through (FT) and bound fractions collected for ESI-LC-MS/MS analyses. Glycopeptides were enriched in the bound fraction as compared to FT.

 

Measuring Plasma Membrane Protein Endocytic Rates by Reversible Biotinylation


JoVE 1669 12/23/2009

University of Massachusetts Medical School

Regulated endocytosis governs the cell surface expression levels of the majority of membrane proteins. Here we utilize reducible, membrane impermeant biotinylation reagents to measure the endocytic rate of the dopamine transporter (DAT), a polytopic membrane protein. The method facilitates a straightforward approach to measuring the endocytic rate of most plasma membrane proteins.

 

In vitro Reconstitution of the Active T. castaneum Telomerase


JoVE 2799 7/14/2011

Gene Expression and Regulation, The Wistar Institute, University of Pennsylvania

Efforts to isolate the catalytic subunit of telomerase, TERT, in sufficient quantities for structural studies, have been met with limited success for more than a decade. Here, we present methods for the isolation of the recombinant Tribolium castaneum TERT (TcTERT) and the reconstitution of the active T. castaneum telomerase ribonucleoprotein (RNP) complex in vitro.

 

Stable Isotopic Profiling of Intermediary Metabolic Flux in Developing and Adult Stage Caenorhabditis elegans


JoVE 2288 2/27/2011

1Department of Pediatrics, The Children's Hospital of Philadelphia, 2Department of Pediatrics, University of Pennsylvania

Stable isotopic profiling by gas chromatography mass spectrometric analysis of intermediary metabolic flux is described in the nematode, Caenorhabditis elegans. Methods are detailed for assessing isotopic enrichment in carbon dioxide, organic acids, and amino acids following isotope exposure either during development on agar plates or during adulthood in liquid culture.

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