Bio-Rad Immun-Star™ AP Chemiluminescence Kits - Manual (Page 17)

Antibody Buffer
(0.2% nonfat dry milk in TTBS)
Add 0.4 g of nonfat dry milk to 200 ml TTBS. Stir until
dissolved. Label this solution “Antibody Buffer”.
Primary Antibody Solution
Dilute the primary antibody to the appropriate titer in 100 ml of
antibody buffer. Label this solution “Primary Antibody
Solution”.
Second Antibody Conjugate Solution (1:3,000)
Add 33 µl of the second antibody conjugate to 100 ml of
antibody buffer. Label this solution “Second Antibody
Solution”.
Substrate Solution for PVDF membrane blots
Immun-Star generates a very fast light signal on PVDF
membrane; therefore, the use of the enhancer is not
necessary. Use 10 ml of chemiluminescent substrate per
200 cm
2 . The substrate is provided ready to use.
Substrate Solution for nitrocellulose membrane blots
Add 500 µl of the enhancer reagent to 10 ml of Immun-Star
chemiluminescent substrate. Label this solution “Substrate
Solution”. This solution can be stored at 4°C for up to 1 week.
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