Alkaline Phosphatase and PBS with Magnesium - (Jul/17/2007 )
Hello,
I am trying to reproduce an ELISA that uses Alk. phosphatase. I'm new to the whole lab environment and while I know theory I don't know much about the practical aspects of lab. Here's what I was wondering.
The protocol I found says that I need to use PBS with Magnesium (though it doesn't say at what concentration). I read that Mg helps the alk. phosph. reaction, but I'm wondering what concentration of magnesium to use and whether the source should be MgCl2 or something else. Any recommendations or specifics would be great. Thank you
-dcotoi-
QUOTE (dcotoi @ Jul 17 2007, 10:59 AM)
Hello,
I am trying to reproduce an ELISA that uses Alk. phosphatase. I'm new to the whole lab environment and while I know theory I don't know much about the practical aspects of lab. Here's what I was wondering.
The protocol I found says that I need to use PBS with Magnesium (though it doesn't say at what concentration). I read that Mg helps the alk. phosph. reaction, but I'm wondering what concentration of magnesium to use and whether the source should be MgCl2 or something else. Any recommendations or specifics would be great. Thank you
I am trying to reproduce an ELISA that uses Alk. phosphatase. I'm new to the whole lab environment and while I know theory I don't know much about the practical aspects of lab. Here's what I was wondering.
The protocol I found says that I need to use PBS with Magnesium (though it doesn't say at what concentration). I read that Mg helps the alk. phosph. reaction, but I'm wondering what concentration of magnesium to use and whether the source should be MgCl2 or something else. Any recommendations or specifics would be great. Thank you
Hi,
usually for most of the enzymes 5mM MgCl2 would be good enough
but not sure about alkaline phosphatase. you could try in google as what is the optimum concentration of Mg for alk. phos.
and it is definite that you have to use it in MgCl2 form
all the best
Leelaram
-leelaram-
i would think that it would be better to use tbs than pbs in the reaction.
-mdfenko-