lab 3

Question 1

what is epitope mapping

Answer 1

looking what eitopes the antibodies bind to

Question 2

what can you do after finding the abs epitope

Answer 2

can mutate Abs to improve affinity

Question 3

what peptides are included

Answer 3

an array of the ones that were injected into the mouse

Question 4

how do you do epepitope mapping

Answer 4

remove blaocking agent form ELISA, wash several times and sopak in washing buffer, take plate then blot dry
add 100ul of dilutent ELISA
add 2ul of sera to each well
incubate for 10-20 mins for mab to bind
flick and add 100ul og secondary Abs that are attcxhaed to horse raddish peroxidase
add 100ul of substrate
stop reaction with sulphuric acid

Question 5

why is dilutent ELISA added

Answer 5

to dilute the sera to make sure it can spread through the well to be able to bind

Question 6

what are the controls used in the ELISA

Answer 6

positive control- mouse monolconal
neg control - no peptide and no monoclonal Ab

Question 7

what colour is positive result

Answer 7

orangey brown insoluble substrate

Question 8

explain the positive result

Answer 8

1 positive result as monocloanl, other wells around will be slightly positive as will hve an overlapping peptode