Help with Pokeweed mitogen

[soe]

In article <451uka$1c0 at tali.UCHSC.edu> ak402 at freenet.HSC.Colorado.EDU (Greg Maniero) writes:
>From: ak402 at freenet.HSC.Colorado.EDU (Greg Maniero)
>Subject: Help with Pokeweed mitogen
>Date: 6 Oct 1995 00:52:26 GMT


>I am beginning to use Pokeweed mitogen in lymphocyte proliferation assays
>and am unsure of storage/handling.  I purchased the PWM from Sigma and
>made a stock sol'n of 1mg/ml in RPMI.  Does anyone know how long this will
>last while refridgerated?  Can I prepare dilutions and freeze for later
>use?  How about thawing and refreezing?   Thanks in advance for any help.

>Greg Maniero
>University of Colorado, Boulder, CO
>Dept. of EPO Biology
>(303) 492-6014

I suggest you should first find out the optimal concentration of PWM which you 
often use. If you use 100 or 200 ul  of 1 million per ml of  cell suspension, 
in each microtitre well then make eleven times concentrated PWM solution and 
kept aliquots (at least 200 ul) in -20 degree centigrade.  The micro 
titertubes (8.8x45mm) are the suitable container for storage. The reagent is 
not expensive approximately US$25 per 5 mg. Do not refreeze.  We check 
the optimal activity of each mitogen every year. Do not know how long they can 
be in the freezer without any deterioration.  I hope it will be of some help.  
Good luck.

Aye Soe   \\\\\                                               
    /////> >>>>>---##########-----    Greg    -----##########---<<<<<>/////    
                                               \\\\\

Aye Soe		                      Internet: SoeA at agresearch.cri.nz
Researcher                               Phone: 64-4-5286089
P.O.Box 40063, Upper Hutt                  Fax: 64-4-5281380
NEW ZEALAND