Home > Community > Is Sodium orthovanadate compatible with RIPA lysis buffer with EDTA?
Upvote

21

Downvote
+ Vanadates
+ Edta
+ Lysis buffer
+ Ripa buffer
Posted by
Mineta Harlow

Is Sodium orthovanadate compatible with RIPA lysis buffer with EDTA?

Don Welty  Follow

I do agree with Dr. ADam, its of no use adding vanadate when EDTA present in the buffer, my suggestion is that try ur experiment without adding vanadate.
all the best

More

Upvote

VOTE

Downvote
Brian McNellis  Follow

''EGTA, on the other hand, forms a weaker complex with vanadate (34) and may therefore be a practical alternative when divalent cations must be chelated''
http://www.jbc.org/content/272/2/843.full

More

Upvote

VOTE

Downvote
Adam Jacholkowski  Follow

It seems that EDTA can bind vanadate*, so it would be pointless to add vanadate as an inhibitor in the presence of EDTA, unless the vanadate concentration is in excess of the EDTA concentration. EDTA itself is a serine/threonine phosphatase inhibitor due to binding Mg2+, which is required for enzyme activity, so vanadate is probably unnecessary anyway.
*http://www.plantphysiol.org/content/75/3/586.full.pdf

More

Upvote

VOTE

Downvote