Preoarations  of K 12 Antisera

0 Antisera

Preparation*

  An overnight slope culture Hartley's Digest Agar was washed into 5 ml.
of saline filtered through cotton wool, washed, and resuspended in saline.
This suspension was then aupclawd at 10 lbs for 20 minutes.
It contained about 54.00 x 10 organisms /ml.

K Antiserum
         --

Pre aration
-%T--
   lope  culture grown and washed of a8 before and resuspended in 5 co
saline.  It wa8 then treated with 25 cc methyl aloohol and shaken vigorously
at intervals for 2 hours.  The alcohol was then centrifuged off and the
deposit washed twice in saline.
of 3700 x lo6 organisms /ml.    The final suspension had a strenghh

H Antiserum

Preparation
-.
   A wellgrown, owrnight broth culture was treated with formalin to make

a 0.2$ solution of this substance.   It is important to use only highly
motile strains for this work.
Strength 4000 x lo6 organi8ms /ml.

Injection Routine

  The same routine was followed with all the above suspensions.
CLt two day interval8 the following doses were given:

0,25 ml.    0.5 ml.    0.5 ml*     1.0 ml. i   1.5 ml.

The rabbits were then bleed f'iw day8 after the last injeotion.