26 LABORATORY COURSE IN SERUM STUDY 



Experiment 1 



To SHOW THAT RED BLOOD CELLS WILL ABSORB AMBOCEPTOR BUT 



NOT COMPLEMENT 



Set up two mixtures in centrifuge tubes as indicated below: 



A. Sheep cells and heated immune rabbit serum 



B. Sheep cells and fresh normal guinea pig serum. 



Incubate for 30 minutes at 37 C., then centrifugalize and remove 

 the supernatant fluid, using a capillary pipette with rubber nipple. 



Wash the sediment once in salt solution. 



Resuspend the washed sediment in each tube in 3 c.c. salt solution. 



Add the reagents as indicated and incubate for one hour at 37 C., 

 observing at short intervals. Tabulate the results. 



Tube A 



Sheep cells 5% 1.0 c.c. 



Inactivated immune rabbit serum l .... 1.0 c.c. 



Incubate 30 minutes at 37 C., centrifugalize, remove the superna- 

 tant fluid to another tube and wash the sediment. 



We now have 2 tubes one (Tube 1) containing the supernatant 

 fluid, the other (Tube 2) the sediment from the original Tube A. To 

 these tubes add reagents as follows : 



* 



Tube 1 Tube 2 



(Supernatant fluid from (Sediment from Tube A 



Tube A) 2.0 c.c. resuspended in salt 



Guinea pig serum (1-10) 1.0 c.c. solution) 3.0 c.c. 



Sheep cells 5% . . . 1.0 c.c. Guinea pig serum (1-10) 1.0 c.c. 



Incubate and observe. 



In which of the two does haemolysis take place? What conclu- 

 sion as to the reaction that has taken place in the original mixture in 

 Tube A can you draw from this? 



TubeB 



Sheep cells 5% 1.0 c.c. 



Guinea pig serum (1-10) 1.0 c.c. 



Incubate 30 minutes at 37 C., centrifugalize, remove supernatant 

 fluid to another tube and wash sediment. 



1 Diluted by the instructor so as to contain two hemolytic units in 1 c.o. 



