Show Advanced Search

REFINE YOUR SEARCH:

Containing Text
- - -
+
Filter by author or institution
GO
Filter by publication date
From:
October, 2006
Until:
Today
Filter by journal section

Filter by science education

 
 
Serum Albumin: A major protein in the Blood. It is important in maintaining the colloidal osmotic pressure and transporting large organic molecules.

Restriction Enzyme Digests

JoVE 5070

Restriction enzymes or endonucleases recognize and cut DNA at a specific sequence. These enzymes occur naturally in bacteria as a defense against bacteriophages - viruses that infect bacteria. Bacterial restriction enzymes cut the invading bacteriophage DNA while leaving the bacterial genomic DNA unharmed due to addition of methyl groups.

This video explains the basic principles of restriction enzymes including: how restriction enzymes are named and the types of recognition sites and overhangs that exist. Also provided is a step-by-step generalized procedure for how to set up a restriction digest including the necessary components, the order in which the mixture should be assembled, and the typical incubation temperature and time. The importance of inactivating restriction enzymes to prevent star activity is mentioned. Tips for performing multiple enzymes digests and using controls in digestion reactions are also provided.


 Basic Methods in Cellular and Molecular Biology

The Western Blot

JoVE 5065

Western Blotting is used to identify the presence of specific proteins in electrophoretically separated samples. Following separation by a technique known as sodium dodecyl sulfate polyacrylamide gel electrophoresis, or SDS-PAGE, western transfer is used to move proteins from a polyacrylamide gel onto a piece of membrane which traps the proteins in their respective locations. Next, the membranes are probed with antibodies in a process called immunboblotting. Immunoblotting uses antibody-protein and antibody-antibody binding through specific recognition sites, providing the high specificity required for identifying a single protein. The detection of antibodies takes place using reporter systems which includes the use of enzymes. Enzymes can be attached to the end of an antibody and react with substrates to produce changes in color or light. These signals can then be imaged and quantified using a process called densitometry. This video-article presents an overview of the western blot technique by describing western transfer, the use of antibody detection, and image analysis. The steps involved with western transfer such as the assembly of the transfer sandwich and transfer conditions are discussed in detail as well as the theory behind antibody binding and detection of those antibodies. The broad applications of this technique are described through severa


 Basic Methods in Cellular and Molecular Biology

Chemically-blocked Antibody Microarray for Multiplexed High-throughput Profiling of Specific Protein Glycosylation in Complex Samples

1Institute for Hepatitis and Virus Research, 2Department of Microbiology and Immunology, Thomas Jefferson University, 3Drexel University College of Medicine, 4Van Andel Research Institute, 5Institute for Hepatitis and Virus Research, Serome Biosciences Inc.

JoVE 3791


 Biology

Imaging of HIV-1 Envelope-induced Virological Synapse and Signaling on Synthetic Lipid Bilayers

1Department of Pathology, New York University Langone School of Medicine, 2Program in Molecular Pathogenesis, Marty and Helen Kimmel Center for Biology and Medicine and Skirball Institute for Biomolecular Medicine, 3Laboratory of Molecular Immunogenetics, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, 4Veteran Affairs New York Harbor Healthcare System

JoVE 3757


 Immunology and Infection

Flow Cytometric Detection of Newly-Formed Breast Cancer Stem Cell-like Cells After Apoptosis Reversal

1School of Life Sciences, Chinese University of Hong Kong, 2State Key Laboratory of Agrobiotechnology, Chinese University of Hong Kong, 3Key Laboratory for Regenerative Medicine, Ministry of Education, Chinese University of Hong Kong, 4Centre for Novel Biomaterials, Chinese University of Hong Kong

Video Coming Soon

JoVE 58642


 JoVE In-Press

Protein Kinase C-delta Inhibitor Peptide Formulation Using Gold Nanoparticles

1Latner Thoracic Surgery Research Laboratories, Toronto General Hospital Research Institute, University Health Network, 2Respiratory Medicine Research Laboratory, Institute of Translation Medicine, Shanghai General Hospital, Shanghai Jiaotong University, 3Institute of Medical Science, Faculty of Medicine, University of Toronto

Video Coming Soon

JoVE 58741


 JoVE In-Press

12345678921
More Results...