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Biochemistry

ImmunoPrecipitation of Nuclear Protein with Antibody Affinity Columns Authors:  Saïda Dadi, Dominique Payet-Bornet and Pierre Ferrier, date: 02/05/2013, view: 20213, Q&A: 1
Co-Immunoprecipitation (Co-IP) is the method used to pull down protein partners of a protein of interest using an antibody that specifically binds to this specific protein in order to test protein-protein interaction. “Pulled down” proteins can be analyzed by western blot for suspected protein partner, or by Mass spectrometry for high throughput ...
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Co-Streptavidin Precipitation Authors:  Saïda Dadi, Dominique Payet-Bornet and Pierre Ferrier, date: 02/05/2013, view: 6972, Q&A: 0
Co-Streptavidin Precipitation (Co-SP) is a method to pull down protein partners of a protein of interest tagged with the streptavidin binding protein domain, and using streptavidin columns that specifically bind to Streptavidin Binding Protein (SBP) in order to test the protein-protein interactions. Proteins of interest to be tested for their ...
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Protein-peptide Interaction by Surface Plasmon Resonance Authors:  Eiji Ishii, Yoko Eguchi and Ryutaro Utsumi, date: 02/05/2013, view: 11787, Q&A: 0
This protocol measures the protein-peptide interaction by surface plasmon resonance (SPR) using Biacore X100 (GE Healthcare). The Biacore system can monitor the direct interaction between biomolecules. There are several methods of immobilizing a ligand to the sensor chip. The optimal immobilization method for each experiment needs to be selected. ...
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Purification of Antigen 85 Complex of Mycobacterium tuberculosis Authors:  Uma Devi Ranganathan and Alamelu Raja, date: 02/05/2013, view: 6171, Q&A: 0
Serodiagnosis of tuberculosis using purified native antigens is one of the approaches for the early detection of TB. The Antigen 85 complex consisting of Ag 85A, 85B and 85C are the most abundant antigens in the culture supernatant of mycobacteria. It has also been shown to be the immunodominant (the property of an antigenic determinant that ...
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Cancer Biology

In vitro Tumorsphere Formation Assays Authors:  Sara Johnson, Hexin Chen and Pang-Kuo Lo, date: 02/05/2013, view: 33784, Q&A: 3
A tumorsphere is a solid, spherical formation developed from the proliferation of one cancer stem/progenitor cell. These tumorspheres (Figure 1a) are easily distinguishable from single or aggregated cells (Figure 1b) as the cells appear to become fused together and individual cells cannot be identified. Cells are grown in serum-free, non-adherent ...
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Isolation of Cancer Epithelial Cells from Mouse Mammary Tumors Authors:  Sara Johnson, Hexin Chen and Pang-Kuo Lo, date: 02/05/2013, view: 8851, Q&A: 0
The isolation of cancer epithelial cells from mouse mammary tumor is accomplished by digestion of the solid tumor. Red blood cells and other contaminates are removed using several washing techniques such that primary epithelial cells can further enriched. This procedure yields primary tumor cells that can be used for in vitro tissue ...
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Developmental Biology

Organotypic Slice Culture of Embryonic Brain Sections Author:  Froylan Calderon de Anda, date: 02/05/2013, view: 10729, Q&A: 0
This technique will allow using brain slices to study several aspects of cortical development (i.e. neurogenesis), as well as neuronal differentiation (i.e. neuronal migration, axon and dendrite formation) in situ. This protocol is suitable for various embryonic stages (Calderon de Anda et al., 2010; Ge et al. ...
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Immunology

In vitro Culture of Human PBMCs Authors:  Santosh K Panda and Balachandran Ravindran, date: 02/05/2013, view: 60379, Q&A: 8
Peripheral blood mononuclear cells (PBMCs) consist of chiefly of lymphocytes and monocytes. Purified PBMCs are used in vitro to evaluate a variety of functions of lymphocytes viz; a) proliferation to mitogenic (PHA, Con-A) stimulation, b) monitoring of prior sensitisation in antigen recall assays by scoring lymphocyte proliferation, c) ...
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Isolation of Human PBMCs Authors:  Santosh K Panda and Balachandran Ravindran, date: 02/05/2013, view: 33086, Q&A: 2
Peripheral blood mononuclear cells (PBMCs) are chiefly lymphocytes and monocytes. PBMCs are separated from the whole blood by a density gradient centrifugation method using Ficoll Histopaque.
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