Bio-Rad Immun-Star™ AP Chemiluminescence Kits - Manual (Page 32)

Tris-buffered saline (TBS)
(20 mM Tris, 500 mM NaCl, pH 7.5)
Add 120 ml of 10x TBS to 1,080 ml of ddH
2
O. Label this
bottle “1x TBS”.
Wash solution (TTBS)
(20 mM Tris, 500 mM NaCl, 0.2% Tween 20, pH 7.5)
Add 2 ml of Tween 20 to 1 L of TBS. Label this solution
“TTBS”.
Avidin-AP or Streptavidin-AP solution
Add 100 µl of avidin-AP or streptavidin-AP to 100 ml TTBS.
Substrate solution for nitrocellulose membrane blots
Add 150 µl of the enhancer reagent to 3 ml of Immun-Star
chemiluminescent substrate. Label this solution “Substrate
Solution”.
For PVDF membrane blots, use the Immun-Star
chemiluminescent substrate without the enhancer reagent.
Total Protein Detection Procedure
Note: Before beginning, read through the entire procedure.
The following procedure is based on 100 ml of each solution,
which is sufficient volume to assay one 10 x 15 cm
membrane.
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