Exam 1 Part 3 Flashcards
parts of antibody
light chain variable region hinge heavy chain constant region
antigen
foreign particle (protein) that the Ab recognizes
Epitope
region/part of the antigen to which the antibody binds
1 antigen can have MANY epitopes
B cells
produce antibodies
each is unique
normal role of antibodies
recognize antigen and clump together so rest of immune system can get rid of it
make antibodies specific to protein of interest
- purify antigent
- find appropriate animal and inject
- animal immune system makes antibodies
- blood draw
polyclonal antibodies
many different epitopes recognized
monoclonal
single epitope recognized
made by combining with tumor cell to create hybrid cell
using antibodies to study cells/proteins
can look at protein expression, isolate protein, means of comparison, structure/function, or target cells in body
immunoprecipitation
when: to isolate your protein of interest
1. conjugate an antibody to a bead
2. add cell lysate to column
3 antibody binds to protein
4. wash column
5. elute off your protein (can change heat, pH, chemicals, etc)
Immunocytochemistry and immunohistochemistry
- label
- fixation/prep
- incubation, wash
- view
amplification of signal
primary antibody not strong enough signal, so secondary antibody that binds to primary constant site added
ELISA
Enzyme-linked immunosorbent assay
antibody sticks to stuff
ELISA direct
put Ab on plate add antigent add another Ab color "is antigent present?'
ELISA indirect
Ag
add sample
2o Ab added with marker
color reaction
