antibodies life - 4oC, -20oC or -80oC (Jun/06/2007 )
If it is better to freeze Ab in aliquots, I wonder why they put the "DO NOT FREEZE" notice in the data sheet.
Since time and experience have proven that Ab is better to be kept frozen, then perhaps I shall do that too..
If one doesnt freeze antibodies, then they might not work in a year or so and then you will have to buy the antibody again which will generate more revenues for the company. Its a trick to make more money.
Or like Santa Cruz uses gelatin in their buffers, so that is why you shouldn't freeze theirs.
If a buffer has BSA or other stabilizer, they are more apt to last longer at 4C than if they are just in PBS.
Very useful information.
Dear all,
Really useful information. But according to an old Worthington Biochemicals catalog description of
peroxidase, sodium azide is an irreversible inhibitor of HRP. Can someone give comment for this topic? Thanks.
"INHIBITION OF PEROXIDASE BY METHANOL AND BY METHANOL-NITROFERRICYANIDE FOR USE IN IMMUNOPEROXIDASE PROCEDURES
WERNER STRAUS 1
1 Department of Biochemistry, The Chicago Medical School, Chicago, Illinois 60612
The cytochemical reaction for peroxidase is partially inhibited by methanol. The addition of a small amount of sodium nitroferricyanide to the absolute methanol causes further inhibition. The activity of peroxidase is completely suppressed by methanol containing 1% sodium nitroferricyanide and l% acetic acid. The antibody to horseradish peroxidase is much less affected by these inhibitors than horseradish peroxidase itself. By using appropriate combinations of these inhibitors and by varying the time or temperature of treatment, the activity of injected horseradish peroxidase, or of endogenous peroxidase in leukocytes, erythrocytes or normoblasts, can be much decreased or suppressed while the specific ability of the antibody to adsorb horseradish peroxidase can be preserved to a considerable extent. KCN and NaN3 inhibit peroxidase activity only moderately under conditions required for immunoperoxidase procedures, i.e., after removal of the inhibitors.
Submitted on June 28, 1971"
If a buffer has BSA or other stabilizer, they are more apt to last longer at 4C than if they are just in PBS.
Very useful information
If a buffer has BSA or other stabilizer, they are more apt to last longer at 4C than if they are just in PBS.
Very useful information
Yes i'm now doing a report so this is a interesting subject to present.
well i added it into primary antibody solution
And for HRP-secondary ones, i think that amersham send them with thimerosal, which acts like NaN3 but guess it's not HRP inhibiting
Hi
how are the storage rules, if ABs are diluted in milk or BSA? Used to keep AB delutions at 4°C for 1-2 months, using it several times without problems (some ABs do better than others)
How about freezing the dilutions at -20°C? Would the ABs still work after thawing them again? Since these are quite expensive, I haven't tried it yet. Does anyone have any experience with this?
Thanks a lot.
Kahlan
hi,
i wud prefer to dilute the antibody in BSA and use for ur experiments.
then add little bit of glycerol (5-10%) and freeze it.
but please check the efficiency before u use this protocl routinely.
gud luk
very useful info.. but i wonder how long can antibody be effective after diluting with milk(working condition) with the sodium azide and stored at -4C for several months........... in my case it have been this way for two months or three and used in cold room overnight..but suddently not working.