Lecture 18 + 19 Flashcards
antibodies
as a group they recognize antigens, but specifically they bind to a single epitope
epitope
a specific spot on a protein
5 varieties of antibodies (immunoglobulins)
igG (main), igM (first), igA (secreted), igD (dispensable), igE (allergies)
gamma globulin (igG)
- monomer
- bivalent
- 80% of serum antibodies
- enhance phagocytosis, neutralize toxins and viruses, protects fetus and newborn
What makes a unique antibody?
random deletions in the light and heavy chains
reinfection
boosts the amount of antibodies produced
secondary antibodies
produced using the tail (Fc) form primary antibodies
polyclonal
several B-cells each make a different antibody that recognizes the antigen; natural response, mixture of antibodies that may bind many proteins/epitopes
monoclonal
single B-cells made immortal so a single epitope is recognized and available for a long time
monoclonal production
b-cells fused with cancerous b-cells to make them immortal; results in hybridoma
western blot
using antibodies to localize a protein that ran through a gel and was immobilized onto a membrane; to determine quantity and band identity on a gel
ELISA
samples placed in wells or spotted onto membranes and protein absorbs to surface; To determine how much of a protein is present in a mixture
agglutination test
To determine the identity of a pathogen
ouchterlony diffusion assay
to determine the antigens/antibodies in a sample
fluorescent staining
to localize proteins in a cell
______ secondary antibodies can bind to a primary
multiple
isotope labelling
125I or 35S attached to protein and as isotope decays the electrons expose x-ray film layered over blot
enzyme labelling
common enzymes like horseradish peroxidase or alkaline phosphatase catalyze a reaction to give a coloured substance or chemiluminescence
fluorescent labelling
a dye is attached that absorbs one colour of light and gives off another
western blot applications
- detection and quantification of a protein from a sample
- expression profiles of a proteinic tissue samples
- verification of a transgenic organism
- detect infections
How to detect infections by western blot?
- by antigens present in organism
- by patient having antibodies against the infection
