Calmodulin, solution, etc. responses

W. Marshall Anderson wanders at IUNHAW1.IUN.INDIANA.EDU
Thu Jan 26 10:33:06 EST 1995


Here are the responses I got to my query on calmodulin
In article <212B3623BC at iunhaw1.iun.indiana.edu>,
wanders at iunhaw1.iun.indiana.edu wrote:

What is the suggested concentration, solution and storage conditions for 
 a stock calmodulin solution?

in article 
<9501060236.AA06261 at quartz.ucs.ualberta.ca>ras-1 at bones.biochem.ualberta.
ca (Colin Rasmussen) wrote:

I would try it in 10 mM Tris, pH 7.5, 1 mM CaCl2, 1 mg/ml calmodulin. It
is very stable and can be kept at 4C, although for long-term storage I
would aliquot it and keep at -70C.

in article <3en08v$c0l at bengal.oxy.edu> akopian at cheshire.cc.oxy.edu 
(Gabriel Akopian) wrote:
I would also like to know what the recommended storage parameters  
are...please either e-mail me directly, or post on this newsgroup

in article <Pine.SOL.3.91.950108111138.18334A at mallard> Brandon S Johnson 
<johnsbs at mail.auburn.edu> wrote: 

I noticed a message in a protein conference from you concerning 
Calmodulin.  I am a graduate student at Auburn University working in 
plant enzymology.  My work has recently tied into the Calmodulin story.  
I am searching for anyone willing to send me a Calmodulin clone.  I 
would like to use the clone, or part of it, to probe a plant cDNA 
library.  I would be happy to supply more details, or anything else for 
that matter.  My work is completely in plant systems, specifically 
Cowpea.  The enzyme I am studying, glutamate decarboxylase, seems to be 
under Calmodulin control.  Perhaps you could point me in some direction 
for help with the Calmodulin.  Thanks for your time, looking forward to 
a reply...

In article <3ep4oe$9ub at mserv1.dl.ac.uk>, LOGAND
<logand at msdos.montpellier.inra.fr> wrote:


 when I was using calmodulin on a regular (almost daily) basis I stored
it at 4 degrees centigrade in 50mM sodium azide.  So, why 4C? Well, I am 
under the impression (ie. somebody told me in the past) that CaM is a 
very stable protein and the main worry would be bugs, so I included the 
sodium azide (this was not a problem since the assay required azide 
anyway). As for concentration, well I think that would have to be 
dependent on the desired final working concentration and the final 
working concentration would depend on the specific
activity of the calmodulin used (commercial CaM comes from a variety of 
sources and with a variety of specific activities therefore quoting only 
a conc. does not give the necessary info for somebody to repeat your 
results). 

in article <ras-1-0801951059260001 at mac03.biochem.ualberta.ca> 
ras-1 at bones.biochem.ualberta.ca (Colin Rasmussen)  wrote:

Azide is okay sometimes, but can interfere with some enzymes (e.g.
anything containing a heme group if I remember correctly), so I would be
judicious in my use of azide.  Calmodulin is very stable, and so we
usually store it in 10 mM Tris pH 7.5, 1 mM CaCl2.  Experience tells us
the calcium helps maintain the stability of the protein.  It is also 
true is can be stored at 4C, although I would have a backup stock in the
freezer for real long-term storage.
W. Marshall Anderson, Ph.D.
Phone:  219 980-6534
FAX:    219 980-6566
E-mail: wanders at indyvax.iupui.edu



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