Exam 1 Part 3

Question 1

parts of antibody

Answer 1

light chain
variable region
hinge
heavy chain
constant region

Question 2

antigen

Answer 2

foreign particle (protein) that the Ab recognizes

Question 3

Epitope

Answer 3

region/part of the antigen to which the antibody binds

1 antigen can have MANY epitopes

Question 4

B cells

Answer 4

produce antibodies

each is unique

Question 5

normal role of antibodies

Answer 5

recognize antigen and clump together so rest of immune system can get rid of it

Question 6

make antibodies specific to protein of interest

Answer 6

1. purify antigent
2. find appropriate animal and inject
3. animal immune system makes antibodies
4. blood draw

Question 7

polyclonal antibodies

Answer 7

many different epitopes recognized

Question 8

monoclonal

Answer 8

single epitope recognized

made by combining with tumor cell to create hybrid cell

Question 9

using antibodies to study cells/proteins

Answer 9

can look at protein expression, isolate protein, means of comparison, structure/function, or target cells in body

Question 10

immunoprecipitation

Answer 10

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)

Question 11

Immunocytochemistry and immunohistochemistry

Answer 11

1. label
2. fixation/prep
3. incubation, wash
4. view

Question 12

amplification of signal

Answer 12

primary antibody not strong enough signal, so secondary antibody that binds to primary constant site added

Question 13

ELISA

Answer 13

Enzyme-linked immunosorbent assay

antibody sticks to stuff

Question 14

ELISA direct

Answer 14

put Ab on plate
add antigent
add another Ab
color
"is antigent present?'

Question 15

ELISA indirect

Answer 15

Ag
add sample
2o Ab added with marker
color reaction

Question 16

flow cytometry

Answer 16

count cells that you have labeled

based on charge

Question 17

Fluorescence-activated cell sorting

Answer 17

FACS. sort cells into tubes

Question 18

Cell breakage

Answer 18

sonication
detergent
homogenizer
pressure cell

Question 19

sonication

Answer 19

high frequency sound waves break open cell

Question 20

detergent

Answer 20

chemical dissolves cell membrane (SDS, trident, etc)

Question 21

Homogenizer

Answer 21

physical way to open cells

Question 22

Pressure cell

Answer 22

force cells through small opening

french press

Question 23

Centrifugation

Answer 23

separate homogenate into fractions

Question 24

Sedimentation coeffienct

Answer 24

Measured in Svedberg units

Question 25

Differential centrifugation

Answer 25

centrifuge at different speeds

Question 26

velocity sedimentation

Answer 26

put homogenate into a solution of sucrose

Question 27

equilibrium sedimentation

Answer 27

density gradient centrifugation | 1 density of sucrose