Page 40 - IBRO_RNA School_Abstract Book

• Incubate in 1:1000 dilution of digoxigenin alkaline phosphatase Fab frag-
ments in 10% FBS in TBST for 4 hours at room temperature.

• Wash with TBST 2-3 times (Can store at 4C overnight)
• Stain using 500 ug/ml NBT & 250 ug/ml BCIP in developing solution
(0.1M NaCl, 0.1M TrisHCl (pH 9.5), 0.05M MgCl2, 0.1% Tween 20 in au-
toclaved DEPC water).

• Watch under a brightfield microscope after 40-45min, if there is a signal
you will observe purple color.
• To stop the above reaction wash it with PBS and add 4% PFA, after this
you can store it for 1-2 days in 4C or image immediately.