Summary
Summary of function practical reagents - Immunology research and clinic (5052MRK6Y)
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Overview of the functions of different reagents used in the practicals
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Overzicht practicaImmunotherapy
Protein buffer maintain pH
MACS buffer for cell isolation applications. Make sure
that cell separations are of highest
purity possible
IL4 and GM-CSF Differentiation of monocytes to dendritic
cells
Biotinylated anti IL12 and IL6 Will bind to IL12 or IL 6 and this can then
be measured. Non-radioactive labeling
HPE buffer Used to dilute the samples
Streptavidin-poly-HRP solution HRP = horseradish peroxidase. Is biotin
binding protein, enabling signal
amplification and detection of
biotinylated antibodies
TMB-substrate solution Is substrate for HRP, a common label
conjugated to antibodies for ELISA. HRP
in combination with hydrogen peroxide
causes oxidation of TMB blue color
Stop solution To stop the reactive. Don’t want this to
continue for to long otherwise all
samples will color. Mostly add this the
moment your blank starts slightly
coloring
Buffy coat = lymphocytes, monocytes and granulocytes
PMBC = T and B lymphocytes, NK cells and monocytes
Amount of cells per ml counted cells * 104 * dilution
Coatings buffer no proteins, because don’t want anything else to stick to the plate
When adding immune cells in ELISA you want many proteins, because you want very
specific binding
phagocytose
Hepes medium Maintains pH despite changes in
carbon dioxide concentration
NBT 1 tablet Diffuses into cells where it is reduced by
reactive oxygen species to produce
formazan crystals, which after dissolution
of DMSO produces a blue color
Complement
Veronal buffer Standard diluent for complement
fixation test
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