Lecture 18 + 19

Question 1

antibodies

Answer 1

as a group they recognize antigens, but specifically they bind to a single epitope

Question 2

epitope

Answer 2

a specific spot on a protein

Question 3

5 varieties of antibodies (immunoglobulins)

Answer 3

igG (main), igM (first), igA (secreted), igD (dispensable), igE (allergies)

Question 4

gamma globulin (igG)

Answer 4

• monomer
• bivalent
• 80% of serum antibodies
• enhance phagocytosis, neutralize toxins and viruses, protects fetus and newborn

Question 5

What makes a unique antibody?

Answer 5

random deletions in the light and heavy chains

Question 6

reinfection

Answer 6

boosts the amount of antibodies produced

Question 7

secondary antibodies

Answer 7

produced using the tail (Fc) form primary antibodies

Question 8

polyclonal

Answer 8

several B-cells each make a different antibody that recognizes the antigen; natural response, mixture of antibodies that may bind many proteins/epitopes

Question 9

monoclonal

Answer 9

single B-cells made immortal so a single epitope is recognized and available for a long time

Question 10

monoclonal production

Answer 10

b-cells fused with cancerous b-cells to make them immortal; results in hybridoma

Question 11

western blot

Answer 11

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

Question 12

ELISA

Answer 12

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

Question 13

agglutination test

Answer 13

To determine the identity of a pathogen

Question 14

ouchterlony diffusion assay

Answer 14

to determine the antigens/antibodies in a sample

Question 15

fluorescent staining

Answer 15

to localize proteins in a cell

Question 16

______ secondary antibodies can bind to a primary

Answer 16

multiple

Question 17

isotope labelling

Answer 17

125I or 35S attached to protein and as isotope decays the electrons expose x-ray film layered over blot

Question 18

enzyme labelling

Answer 18

common enzymes like horseradish peroxidase or alkaline phosphatase catalyze a reaction to give a coloured substance or chemiluminescence

Question 19

fluorescent labelling

Answer 19

a dye is attached that absorbs one colour of light and gives off another

Question 20

western blot applications

Answer 20

• detection and quantification of a protein from a sample
• expression profiles of a proteinic tissue samples
• verification of a transgenic organism
• detect infections

Question 21

How to detect infections by western blot?

Answer 21

• by antigens present in organism

- by patient having antibodies against the infection