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Media
1. PZ = 9 g NaCl + 1000 Aqd (Aquades)
2. Prog-11-BSA-Sol = 100 ug P4-11-BSA +
1 ml PZ
3. P4-11-BSA Emulsion = 0.5 sol P4-11BSA + 0.5 ml FCA
Mix well by using 2 syringe that
connected each other with small canule,
pumping several times emulsion?
Tester (??)
Why should apply FCA?
There are containing with oil, the mechanism?
There are also with weaked mycobacterium,
what the function and mechanism?
How to collect blood serum in rabbit
Clean up one ear
A small incision at V marginale
The ear is entered into graded
Erlenmeyer glass connected with canule
and vacuum machine
The blood will come up smoothly and
stops it after 15 20 ml
The blood let clotting for 2 3 hours
One hour after collected, release the
cloth by puncturing at glass wall
Another 1 2 hours later the serum will
come out proportionally, max 60% of
blood can be harvested as NRS.
Immunitation
Intradermal Injection
Clinical Response
1st response
Laten period there take 5 7 D
No any Ab increase
Body temp arise
2. Primary Ab response 10 30 D, just
immature response of increasing Ab,
mostly due to of B-cell response
3. 2nd response, a optimum to mas
response of increasing Ab mostly by Tcell
2nd response, (following booster)
Result
X cpm A/S x cpm NRS
% B = -------------------------------- x 100%
Titration of A/S
Media PBS:
A. 1.42 g Na2HPO4 1000 cc aquades
B. 1.38 g Na2HPO4.H2O 1000 cc
aquades
Take 600 ml A + 400 ml B mix
Put in 9 g NaCl and 1 g sod azide
A part (500 ml) of this sol add with 500
mg gelatin
NRS serial dilution (1 10)
1:50, 1:100 1:25600 (2 x each Step
Lower)
A/S serial dilution (110)
1:50, 1:100 1:25600 (2 x each Step
Lower)
Protein A-sol
P4-11-succinyl125I diluted 20.000
CPM/ minute (gamma counter)
Cold
PBS
0
0
3
500
200
100
100
500
200
100
100
500
200
100
100
PBSG Diluted
(ul)
SRA
(ul)
500
200
100
500
200
100
100
500
200
100
100
500
200
100
100
And soon
Tube
NRS
1:25600
NRS
500
1:25600
A/S
500
1:25600
Prot Cold
A
PBS
(ul)
100 3
200
100
100
200
100
100