TEACHER

Pre-lab Questions Answer Key

1. What is an antigen?
Any agent that provokes an acquired immune response and is bound specifically by
either antibodies or T cells. Binding occurs at a specific region called an epitope on
the antigen. An antigen may have one or more epitopes.
2. What is an antibody?
Immunoglobulin protein formed in response to a challenge of the immune system by
a foreign agent (antigen).
3. How do antigens and antibodies interact?
An antibody binds to a specific antigen at a specific epitope.
4. What does !"#A stand $or?
Enyme!linked immunosorbent assay
%. Why do you need to assay &ositi'e and negati'e control sa(&les as well as your
e)&eri(ental sa(&les?
"ontrols are needed to make sure the assay is #orking correctly. If there are no
positive controls and the sample is negative$ #e can%t kno# if the sample #as truly
negative or if assay didn%t #ork. "onversely$ #ithout a negative control$ there is no
#ay of kno#ing if all samples (positive or not) #ould have given a positive result.
*. Why did the county health de&art(ent want to test both the dead &igeon as well
as +ar$ield?
The disease has been kno#n to infect cats as #ell as birds. &ou #ant to be sure that
'arfield is going to be around for a #hile longer(
Analysis Questions Answer Key
,. -id your seru( sa(&le contain the antibody against H%? How do you .now?
)ifty percent of the samples (bird samples) should be * for the antibody and +,-
should be . ('arfield samples). If the sample is positive$ there should be a color
change. (blue)
/var/###/apps/conversion/tmp/scratch01/22345,+63.doc 1/24/2,1, 15741 A1/81 1
/. Why did you add a &ri(ary antibody?
The primary antibody has a specific shape that matches an epitope on the antigen. If
the serum sample #as positive for the antibody$ the primary antibody in the bird
serum #ould bind to the 9+ antigen originally placed on the plate. If the serum
sample #as negative$ the primary antibody did not bind and #as flushed out in the
#ash step.
0. Why did you add a secondary antibody?
The secondary antibody is covalently bound (con:ugated) to an enyme that catalyes
a chemical reaction #hen the enyme substrate is added. This #ould produce a color
change if the serum sample contained antibody to the 9+$ because the enyme bound
secondary antibody #ould bind to the primary antibody already bound to antigen in
the #ells. (This occurs because of the specificity of the secondary antibody to the
shape of the primary antibody.) If the serum test sample did not contain 9+ antibody$
(primary antibody) there #ould have been no binding in the #ells$ so the secondary
antibody #ould have nothing to bind to and #ould be flushed out in the #ash step.
11. Why did you assay your sa(&les in tri&licate?
Assaying the samples in triplicate is another control. If you do not get the same result
in all triplicate #ells$ you have a problem #ith your e;perimental technique or you
have made a pipetting error. In a clinical laboratory$ the e;periment #ould have to be
repeated.
11. What antibody-based tests can you buy at your local &har(acy?
Test kits that are based on the same principles as the E<I=A include home pregnancy
and ovulation tests and tests for the presence of illegal drugs such as mari:uana and
cocaine.
/var/###/apps/conversion/tmp/scratch01/22345,+63.doc 1/24/2,1, 15741 A1/81 2