AMBOCEPTORS AND COMPLEMENTS 927 



separate serum, and dilute 1 : 20 with normal salt solution. Secure antisheep and 

 antihuman hemolytic serums (inactivated) whose hemolytic titers are known. 



2. Proceed as follows: 



Tube 1: 1 c.c. sheep corpuscles+1 c.c. complement (l:20)+two doses of anti- 

 sheep hemolysin + salt solution up to 3 c.c. 



Tube 2: 1 c.c. sheep corpuscles+1 c.c. complement (l:20)+five doses of anti- 

 human hemolysin. 



Tube 3: 1 c.c. human corpuscles+1 c.c. complement (l:20)+two doses of anti- 

 human hemolysin. 



Tube 4: 1 c.c. human corpuscles+1 c.c. complement (l:20)+five doses of anti- 

 sheep hemolysin. 



Tube 5: 1 c.c. sheep corpuscles +2 c.c. salt solution (control). 



Tube 6: 1 c.c. human corpuscles+1 c.c. salt solution (control). 



3. Shake tubes gently and incubate for an hour or two. 



(a) In which tubes has hemolysis occurred? 



(b) What does this experiment teach as to the specificity of these 

 amboceptors? 



(c) Discuss the specificity of hemolytic and bacteriolytic ambo- 

 ceptors. 



EXPERIMENT 77. GENERAL PROPERTIES OF AMBOCEPTORS 



1. Secure antisheep amboceptor so diluted that 1 c.c. contains 1 hemolytic unit. 



2. Prepare a 2 per cent, suspension of sheep corpuscles. 



3. Secure fresh guinea-pig complement serum and dilute 1 : 20. 



4. Place two units of amboceptor into each of six test-tubes. Place these in a 

 water-bath and heat at 60 C. 



5. Remove a tube from the water-bath after five minutes, ten minutes, fifteen 

 minutes, thirty minutes, forty-five minutes, and one hour. After allowing them 

 to cool, add 1 c.c. corpuscle suspension and 1 c.c. complement serum (1 : 20). 



6. Set up a tube containing two units of amboceptor (not heated) + 1 c.c. cor- 

 puscle suspension +1 c.c. complement serum (control). Shake each tube gently and 

 incubate for an hour at 37 C. 



(a) Has the amboceptor deteriorated by exposure to this degree of 

 heat? 



(b) Discuss the stability of amboceptors to temperature, age, germi- 

 cides, and drying. 



(c) Discuss the relative stability of amboceptors and complements. 



EXERCISE 30. AMBOCEPTORS AND COMPLEMENTS. HEMOLYSINS 



(Continued) 



EXPERIMENT 78. MECHANISM OF AMBOCEPTOR ACTION 



1. Secure antisheep amboceptor so diluted that 1 c.c. contains one hemolytic 

 unit. 



2. Prepare a 2^ per cent, suspension of sheep corpuscles. 



3. Secure fresh guinea-pig serum and dilute 1:20. 



