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Author Topic: Beta-galactosidase ABC based protocol ??  (Read 5957 times)

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Offline drcarter

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Beta-galactosidase ABC based protocol ??
« on: February 15, 2005, 03:46:41 AM »
Hi,

I would like to do an IHC with a beta-galactosidase ABC based method ie avidin/streptavidin + biotinylated beta-galactosidase.
Do you know any commercial kit available? I haven't found any.

If I have to to the ABC complex by myself, what amount of avidin/streptavidin should I mix with biotinylated beta-galactosidase in order it works well ?

Thanks for your help.

Beta-galactosidase ABC based protocol ??
« on: February 15, 2005, 03:46:41 AM »

Offline angela

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Beta-galactosidase ABC based protocol ??
« Reply #1 on: February 15, 2005, 05:00:40 PM »
Hi,
I guess you can also use a biotinylated secondary antibody and then the ABC kit. That is what I have done and it works fine. My secondary was a Pharmingen, and the ABC is a Standard Vector kit.
hope this can help
Angela

Offline richard03

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Beta-galactosidase ABC based protocol ??
« Reply #2 on: February 15, 2005, 10:29:31 PM »
Since your primary is biotinylated beta-galactosidase, you can directly use HRP-streptavidin (1:500, Vector Labs) as detection agent.

Richard

Offline drcarter

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Beta-galactosidase ABC based protocol ??
« Reply #3 on: February 28, 2005, 05:14:13 AM »
Thank you for your suggestion.
I am finally going to use Betagal-streptavidin method directly on biotynilated secondary Ab, purchased from Roche, as I have read that LSAB method is better than ABC based one.
drcarter

Beta-galactosidase ABC based protocol ??
« Reply #3 on: February 28, 2005, 05:14:13 AM »