Western blot problem, need suggestions

Igor josstalin at home.com
Fri Jul 20 00:16:59 EST 2001


NBT/BCIP is for alkaline phosphatase (AP), not for horse redish
peroxidase (HRP)

oddie wrote:
> 
> 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.
> ;)
> 
> <http://www.biowww.net/forum/read.php?f=1&i=2880&t=2880>




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