Transcript BIO132 Lab 5 Exercise 35A/35 Lymphatic System Fig. 35A.1/35.1

BIO132 Lab 5
Exercise 35A/35
Lymphatic System
Fig. 35A.1/35.1
Fig. 35A.2 in 8th & 9th editions
Figure 35A.3 in 10th edition
Figure 35.3 in 11th & 12th editions
Lymphatic Vessel
4X Objective
(40X)
10X Objective
(100X)
Valve
Czura 2005
4X Objective
(40X)
Lymph Node
Follicle
Medulla
Cortex
Capsule
10X Objective
(100X)
40X Objective
(400X)
Follicle
Germinal Center
Czura 2005
4X objective
(40X)
Spleen
10X objective
(100X)
40X objective
(400X)
Red Pulp
Lymphocytes
White pulp
Czura 2005
4X Objective
(40X)
Palatine Tonsil
Crypt
Crypt
Follicle
10X Objective
(100X)
40X Objective
(400X)
Follicle
Germinal center
Czura 2005
Activity 3: Ouchterlony Test
-allows visualization of antibody-antigen reaction
-detects the presence of particular antigens in extracts
-antigens and antibodies are placed in wells in a gel and
allowed to diffuse toward each other
-if an antigen reacts with an antibody a precipitin line forms
-precipitin line = agglutination of antigen by antibodies
Ab
Ag
Figure 35A.6 in 8th & 9th
Fig. 35A.8 in 10th
Fig. 35.8 in 11th & 12th
I
1
3
4
1
III
2
2
4
2
1
3
II
Section I
1
Red dye in well
2
Green dye in well
Section I
1
2
Diffusion after one hour
Section II
Fill wells with indicated
solutions:
 = “anti”, indicates an antibody
against the named antigen
Horse serum albumin
-horse albumin
3
 -bovine albumin
1
2
4
 -swine albumin
Horse serum albumin is the test antigen
Assay using three different antibodies: one
against horse albumin, one against cow
albumin and one against pig albumin
Section II
16 hours later...
3
1
2
4
Horse serum albumin
-horse albumin
 -bovine albumin
 -swine albumin
Precipitin line forms
where antibody
binds to antigen.
Antibodies are
specific for one
antigen and react
with only that
antigen.
A real plate would look like 
Draw this result!
3
1
http://www.snv.jussieu.fr/bmedia/ATP/images/ouchb1.jpg
4
2
Section II
Section III
Draw this result!
3
1
2
4
Refer to table for well contents: 35A.2 / 35.2 in editions 8-11
35.3 in 12th edition
Section III
Draw this result!
3
1
4
2
What is the unknown antigen?
Refer to table for well contents: 35A.2 / 35.2 in editions 8-11
35.3 in 12th edition
Section III
Draw this result!
-bovine albumin
3
1
2
4 -swine albumin
-horse albumin
What is the unknown antigen?
Bovine (cow) albumin