Documente Academic
Documente Profesional
Documente Cultură
Presented by
Vaibhav Jain
Department of Biophysics
National Institute of Mental Health and Neurosciences
Overview
Immunological techniques include both experimental methods to study
the immune system and methods to generate or use immunological
reagents as experimental tools.
Immunoprecipitation
Immunoelectrophoresis
Immunostaining
Immunoblotting
Flow Cytometry
Sensitivity of various Immunotechniques
Immunoprecipitation
Immunoelectrophoresis
Immunoelectrophoresis combines
electrophoresis with immunodiffusion.
The ELISPOT assay can be used to determine the number of cells within a
sample that are secreting a particular cytokine.
Western Blotting
Principle:
Western blotting identifies and
provides preliminary quantitation of a
specific protein in a complex mixture of
proteins.
Sigma-Aldrich
Gel electrophoresis separates proteins by size,
so Western blot not only provides quantitation,
but also provides information about the
molecular weight of antigen.
Thermo Fisher
Fluorescence activated Cell sorting (FACS)
Research Applications
Multiplexing immunoassays
Multiparameter immunophenotyping
Measurement of intracellular cytokines
Signal transduction pathways
Cell cycle analysis
Measuring cellular function
Immunostaining
Antibodies can be used to
visualize the presence of antigen
within the cell or the tissue.
In immunofluorescence the
fluorescence labeled antibody is
incubated with the cells or tissue
Figure: Antibody stained rat brain
and are visualized under the (including the ventricles and the CA3
region of the hippocampus) tissue.
fluorescence microscope.
Cell Cycle analysis
Bromodeoxyuridine incorporates into DNA
in place of deoxythymidine during DNA
synthesis.