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Reagents
PBS
(0.01M):
Volume:
1 liter,
Na 2HPO 4-------------------- 1.09 g
NaH 2PO 4-------------------- 0.32 g
NaCl ----------------------------- 9 g
Distilled water ------------- 1000 ml
Mix to dissolve and adjust pH to 7.4
4%
Paraformaldehyde in PBS:
Paraformaldehyde
------------------ 40 g
0.1M Phosphate buffer ------------ 1000 ml
Heat
PBS to 60-65 C while stirring. When 60 C is reached, add
paraformaldehyde slowly to stirring PBS solution (Adding
a few drops of 1N NaOH helps the solution clear). Continue
to stir to dissolve the paraformaldehyde, making sure that
the temperature stays between 60-65 C. Cool the solution
untill solution becomes clear and filter and let cool.
4%
Paraformaldehyde-1% Glutaraldehyde in 0.1M PB:
Paraformaldehyde
-------------- 40 g
0.1M Phosphate buffer -------- 1000 ml
Heat to 60-65 C while stirring. Add a few drops of 1N NaOH
until solution clear. Continue to stir to dissolve. Cool
the solution and filter. Add 20 ml of 50% glutaraldehyde
and mix well.
Antigen
Retrieval Solution:
10mM
Sodium Citrate Buffer:
To prepare 1000 ml,
Sodium citrate -------------- 2.94 g
Distilled water ------------- 1000 ml
Adjust pH to 6.0
1
N HCl:
To
prepare 100 ml,
10N HCl --------------------- 10 ml
Distilled water -------------- 90 ml
Mix well.
Blocking
Solution:
2%
Normal Rabbit Serum in PBS:
To prepare 100 ml
Normal rabbit serum ----------- 2 ml
PBS ----------------------------- 98 ml
Mix to dissolve.
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