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Immuno-fluorescence Assay of Leptospiral Surface-exposed Proteins
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1Department of Medicine, David Geffen School of Medicine,University of California, Los Angeles, 2Research service, 151,Veterans Affairs Greater Los Angeles Healthcare System, 3Departments of Medicine, Urology at David Geffen School of Medicine and Department of Microbiology, Immunology and Molecular Gentics,University of California Los Angeles (UCLA), 4Division of Infectious Diseases, 111F,Veterans Affairs Greater Los Angeles Health Care System
Chapters
- 00:05Title
- 02:20Fixation of Leptospira to Glass Slides
- 04:51Labeling with Specific Antibodies
- 06:33Visualization of Leptospires
- 08:42Data Analysis and Representative Results
- 10:20Conclusion
An efficient method to assess surface-exposure of leptospiral proteins is described. The method is specifically designed to avoid disruption of the fragile outer membrane of leptospiral cells. This technique requires employment of several negative controls to assess the integrity of the outer membrane and specificity of antibody reaction.
Tags
Immuno-fluorescence AssayLeptospiral Surface-exposed ProteinsBacterial Surface ProteinsCellular LocalizationFunctional RoleVirulence FactorsPathogenicity MechanismsFractionating Leptospiral MembranesOuter-membrane Proteins (OMPs)LipoproteinsTransmembrane OMPsHydrophobic InteractionLipid Bilayer PhospholipidsAmphipathic β-sheetsBarrel-like StructureOuter Membrane IntegrityImmunofluorescence Assay (IFA)
