lab techniques and immunological disorders Flashcards
what are the 3 areas that can be found within antibody/antigen mixtures
antibody excess, equivalence point, antigen excess
what is the antibody excess
The concentration of antibody exceeds that of the antigen
what is the equivalence point
The concentration of antibody is equal to that of the antigen
what is antigen excess
The concentration of antigen exceeds that of the antibody
what are the main techniques for studying immunology and haematology
- agarose gel electrophoresis
- ELISA
- western blot
- immunocytochemistry
- immunohistochemistry
what is gel electrophoresis
the separation by the movement of charged molecules in an applied electrical field (most commonly in the pcr test for DNA )
do smaller or larger molecules move further down the electrophoresis plate
smaller
why do the DNA fragments move down a gel electrophoresis plate
because they’re negatively charged
what does the movement of the molecules depend on
size, shape, electrical field strength
why is a buffer added to the gel electrophoresis
to provide electrolytes to facilitate the movement of current
what is the gel for in a gel electrophoresis
- retards the movement of molecules according to their dimensions
what is the loading end of a gel electrophoresis plate
the side where the wells are
what does ELISA stand for
enzyme linked immunosorbent assay
what are the main steps of the ELISA test
1 - well is pre-coated with capture antibody
2 - antigens are introduced, including the one of interest
3 - antigen binds to capture antibody
4 - enzyme-labelled detection antibody binds to analyte
5 - the interaction of substrate and enzyme creates a colour change
what is immunohistochemistry and immunocytochemistry use for
its used to determine the anatomical location of an antigen
how do immunocyto and immunohistochemistry work
-a frozen sample of tissue is frozen with a labelled antibody (fluorescent probe)
- the bound antigen is then located using microscopy
- this gives a visual image
what are the 2 types of auto immune disease
organ specific and non-specific (systemic)
what is organ specific auto immune disease
where antibodies and T cells react to self antigens localised in a specific tissue
what’s an example of organ specific autoimmune disease and what organ does it effect
narcolepsy - the brain/ neurological system
what’s an example of a non-specific autoimmune disease
lupus
how do auto immune diseases develop
they’re developed due to genetic susceptibility and environmental triggers
how can virus’s trigger an auto immune disease
if the virus mimics the shape of the bodies antibodies it can cause the body to attack itself as it cannot separate the virus to the bodys own antigen
what is gestational diabetes mellitus (GDM)
when a mother becomes diabetic during pregnancy due to a lack of insulin production and an excess of free glucose
what are the maternal impacts of GDM
hypertension (high BP), preterm labour
