1) ELISA - Enzyme-Linked Immunosorbent Assay
i. What is the purpose of an ELISA?
- The purpose of ELISA is to immobilize certain antigens and incubated them with antiserum so that the concentration of the antibody in the antiserum is quantified using an enzyme-linked anti-immunoglobulin antibody also known as an ELISA.
ii. Explain in your own words the following types of ELISAs: direct, indirect, capture or “sandwich”?
- Direct ELISA: The direct ELISA is a more simple method, in addition to it being faster than other ELISA methods. It requires fewer steps and just one antibody to be able to detect the enzyme attached to the bound antibody through a substrate. This substrate is what causes a color change, fluorescence or luminescence. …show more content…
Separate primary antibodies from different species are used along with a secondary antibody from a different species from which the primary antibody was extracted in order to create fluorescence in different parts of the cells, such as the nucleus or the mitochondrion. The purpose of Immunohistochemical staining can evaluate the presence and location of proteins in tissue sections. Immunohistochemical staining enables the observation of processes in the context of intact tissue which is very useful in the ability to assess the progression of disease such as cancer. Some disadvantages are that the procedure is less sensitive quantitatively than immunoassays, such as western blotting or …show more content…
What is the purpose of Flow Cytometry?
- The purpose of Flow Cytometry is to be able to analyze the expression of the cell surface along with intracellular molecules and being able to characterize and define different cell types in heterogeneous cell populations as well as being able to analyze cell size and volume. It allows for simultaneous multi-parameter analysis of surface molecules on single cells and more advanced cytometers can also sort cells based on their parameters. A disadvantage is that this method is complex, expensive and multiple controls are needed for multicolor
i. What is the purpose of an ELISA?
- The purpose of ELISA is to immobilize certain antigens and incubated them with antiserum so that the concentration of the antibody in the antiserum is quantified using an enzyme-linked anti-immunoglobulin antibody also known as an ELISA.
ii. Explain in your own words the following types of ELISAs: direct, indirect, capture or “sandwich”?
- Direct ELISA: The direct ELISA is a more simple method, in addition to it being faster than other ELISA methods. It requires fewer steps and just one antibody to be able to detect the enzyme attached to the bound antibody through a substrate. This substrate is what causes a color change, fluorescence or luminescence. …show more content…
Separate primary antibodies from different species are used along with a secondary antibody from a different species from which the primary antibody was extracted in order to create fluorescence in different parts of the cells, such as the nucleus or the mitochondrion. The purpose of Immunohistochemical staining can evaluate the presence and location of proteins in tissue sections. Immunohistochemical staining enables the observation of processes in the context of intact tissue which is very useful in the ability to assess the progression of disease such as cancer. Some disadvantages are that the procedure is less sensitive quantitatively than immunoassays, such as western blotting or …show more content…
What is the purpose of Flow Cytometry?
- The purpose of Flow Cytometry is to be able to analyze the expression of the cell surface along with intracellular molecules and being able to characterize and define different cell types in heterogeneous cell populations as well as being able to analyze cell size and volume. It allows for simultaneous multi-parameter analysis of surface molecules on single cells and more advanced cytometers can also sort cells based on their parameters. A disadvantage is that this method is complex, expensive and multiple controls are needed for multicolor