ELISA SUBSTRATE BUFFER
(E.S.B.)
0.05M Sodium Phosphate
0.02M Citric Acid
MAKE UP
0.2M Sodium Phosphate
Na2HPO4 FW142.00
28.4g/liter
(0.2M = 4X)
(0.2M
divided 0.05M=4(X))
(1000ml
(1L) divided 4 = 250mls) 250mls
0.1M Citric Acid
(HOC(COOH)(CH2COOH)2.H2O FW210.1
21.01g/liter
(0.1M = 5X)
(0.1M divided 0.02M = 5(X))
(1000ml
(1L) divided 5 = 200mls) 200mls
distilled H2O 550mls
final total 1000mls (1L)
Add 500ml d H2O only
Check pH ( pH should be ph5.0 to 5.13) bring the soln to correct pH use only
0.2M Sodium Phosphate (BASE) brings it up
0.1M Citric Acid (ACID)
brings it down
ON ALL BUFFERS MADED THE LABEL MUST HAVE
DATE/CLASSIFICATION LETTER/ INITIALS