Chemiluminescent substrate for alk-phos?

Don A. Walthers don at meibm4.cen.uiuc.edu
Wed Jul 19 15:07:48 EST 1995


In <3ugnni$4mj at bubba.NMSU.Edu> dkim at nmsu.edu (Daniel Kim) writes:


>Hello:

>I am using alkaline phosphatase conjugated second antibody system for 
>Western blot detection.  I have been dissatisfied with the density of 
>signal and photoreproducibility of NBT/BCIP color signal, and would like 
>to try out a chemiluminescent system.

>Is there a chemiluminescent substrate for alk phos that I could use in 
>place of the NBT/BCIP system I am using now?  If possible, I would prefer 
>to buy and prepare my own reagent solutions, for the sake of expenses, 
>but would accept a kit recommendation if necessary.

>Daniel Kim
>dkim at nmsu.edu

we're using cdp-star from boehringer mannheim. they give you a recipe 
for the detection buffer which is used to dilute the substrate so no
kit is needed. the incubation is only 5min, the signal is detectable 
immediately and stays level for about a day. exposure times are in
the seconds to minutes range. we use a dry immunostaining method where
the PVDF membrane is not rewetted. this may contribute to the clean 
signal we get.

-- 
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|Don Walthers  | don at uiuc.edu  | University of Illinois at UC - Microbiology |
|http://www.cen.uiuc.edu/~don/ | Engineering Workstations - Site Operator    |
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