NBT/BCIP is for alkaline phosphatase (AP), not for horse redish
>> Dear all, I did western blot for three times but I didn't get any positive
> signal. I use 10 % acrylamide gel in order to purifyand detect the ~130 kDa
> Cry toxin receptor protein by western blot. I used cry4B toxin as first
> anitbody(10 ug/ml), antibodies against Cry toxin as second anitbody (1:2000)
> and anti-rabbit AB labeled with HRP as third antibody(1:7000). But I didn't
> get any signal after added BCIP/NBT. My friend told me maybe came from the
> platform shaker, ~ 70 rpm. What should I do ? Thank you for all suggestions.