RE: [Histonet] Storage of Primary Antibodies!

From:Gayle Callis

I generally store antibodies at -27C, never colder, probably more due to
ready freezer space. My immunologist who develops monoclonals/polyclonals
also stores his in house antibodies at this temperature, but NEVER is a
self defrosting freezer.  If the antibody is an inhouse supernate, we add 1
ul 1M sodium azide per ml of supernate.  Aliquots are made of supernates,
and in general we don't bother to purify or concentrate these, they are
totally clean.  We tend to use these as a supernate, often never diluted
further.  We have also had supernates with azide added stay potent for
YEARS at 4C temps, as long as they are stored in sterile tubes, and
aliquoted under sterile conditions in a biohood.  The amount of azide is
not terribly significant with HRP since so many reagents and rinses take
care of getting rid of it before HRP step.  We are careful to use good
tight screw top tubes, have had problems with flip tops - evaporation was a

I think that some antibodies are fussy about storage, per this discussion. =20

At 09:41 AM 2/16/2004 -0500, you wrote:
>        Hi Pam,
>                Thank you for the timely reply. This particular antibody 
>is an "in-house" antibody so I cannot readily give out the clone 
>information. My reasoning for aliquoting and freezing was my concern that=20
>there are no preservatives (like sodium azide) added. However, I conducted=20
>one titration experiment with beautiful results. Then it was about 2 1/2 
>to 3 weeks later until I could conduct the complete experiment with all 
>time points included. At this time I started having problems with the 
>primary antibody dilutions. I suspect that the antibody does not like 
>being frozen. I am using the same procedure and all the same reagents. 
>Jennifer Hoover
>BI Research
>Eli Lilly and Company
>Pamela Marcum 
>02/16/2004 09:22 AM
>        To:     Jennifer Hoover 
>        cc: 
>        Subject:        RE: [Histonet] Storage of Primary Antibodies!
>Hi Jennifer,
>Some antibodies are fine for storage at -80 and some are not.  I think 
>will depend on the antibody.  It may be useful to let us now which
>antibodies are not doing well as other people may have had experience with
>them and have suggestions for you.
>Pam Marcum 
>> -----Original Message-----
>> From:
>> []On Behalf Of Jennifer
>> Hoover
>> Sent: Monday, February 16, 2004 8:48 AM
>> To:
>> Subject: [Histonet] Storage of Primary Antibodies!
>>         Histonet,
>>                 I have a question regarding the storage (short or longer
>> term) of primary antibodies. Is it conducive to aliquot and store 
>> antibodies at -80C?? Is there the potential for diminished activity 
>> storing the antibody for several weeks to several months? The aliquots
>> that were used were thawed and used for one day only. Any volume that 
>> not used was disgarded so freeze/thawing is not the problem. I would
>> welcome any insight. Thank you very  much!
>> Jennifer Hoover
>> BI Research
>> Eli Lilly and Company
>> _______________________________________________
>> Histonet mailing list
>Histonet mailing list
Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology 
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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