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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Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag


JoVE 3370 1/16/2012

School of Biotechnology, Department of Proteomics, Royal Institute of Technology

A novel and highly efficient two-step affinity chromatography protocol has been developed and is described in detail. The method is based on a small purification tag with two inherent affinities and is applicable to a wide range of target proteins with different properties.

 

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.

 

Human Pancreatic Islet Isolation: Part II: Purification and Culture of Human Islets


JoVE 1343 5/26/2009

Department of Surgery, University of Illinois, Chicago

Achieving high quality and appropriate quantity of human islets is one of the prominent prerequisites for successful islet transplantation. In this video, we describe step by step the procedures for human pancreatic islet isolation (part II: purification and culture of human islets) using a modified automated method.

 

Streamlined Purification of Plasmid DNA From Prokaryotic Cultures


JoVE 2407 1/05/2011

Pall Life Sciences

This protocol is a cost effective alternative for efficient parallel clarification and plasmid DNA purification from E. coli cultures. The AcroPrep Advance process starts with an optimized lysate clarification filter plate followed by purification on a high binding capacity DNA binding filter plate.

 

Extracting DNA from the Gut Microbes of the Termite (Zootermopsis Angusticollis) and Visualizing Gut Microbes


JoVE 195 5/28/2007

Department of Environmental Science and Engineering, California Institute of Technology - Caltech

This video illustrates the technique for extracting DNA from the species of microbes resident in the termite hindgut. The preparation of a wet mount slide, which is useful for visualizing the gut microbial community is also illustrated, and a tour through the species-rich gut environment is given.

 

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.

 

Expansion, Purification, and Functional Assessment of Human Peripheral Blood NK Cells


JoVE 2540 2/02/2011

Division of Pediatrics, MD Anderson Cancer Center - University of Texas

Here we describe a method to efficiently expand and purify large numbers of human NK cells and assess their function.

 

Identification of Protein Interacting Partners Using Tandem Affinity Purification


JoVE 3643 2/25/2012

Section of Virology, Department of Medicine, Imperial College London

Tandem affinity purification is a robust approach for the identification of protein binding partners. As proof of concept, this methodology was applied to the well-characterized translation initiation factor eIF4E to co-precipitate the host cell factors involved in translation initiation. This method is easily adapted to any cellular or viral protein.

 

Purification of Hsp104, a Protein Disaggregase


JoVE 3190 9/30/2011

Department of Biochemistry and Biophysics, University of Pennsylvania

Here, we describe a protocol for the purification of highly active Hsp104, a hexameric AAA+ protein from yeast, which couples ATP hydrolysis to protein disaggregation. This scheme exploits a His6-tagged construct for affinity purification from E. coli followed by anion-exchange chromatography, His6-tag removal with TEV protease, and size-exclusion chromatography.

 

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.

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