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Antibody Storage
Antibody Storage and Handling
Hevner Laboratory
University of Washington School of Medicine
1. General principles
a.
Antibodies are precious resources
and must be conserved—i.e.,
use as little as possible, do not waste, and keep cold.
b. Antibodies are proteins
and should be kept cold
(refrigerated, on ice, or frozen) when not in use.
c. The
more dilute the
antibody is, the less stable.
Therefore, store antibodies in concentrated form without dilution.
d. Companies that supply
antibodies generally also supply instructions on storage and
handling. Read and follow the
instructions.
e. Antibodies work best when freshly diluted. Do
not dilute your antibodies to make working solutions for
immunostaining until you’re ready to start the incubations.
f.
Repeated freezing and thawing kills antibodies.
Once you have thawed an antibody solution, thereafter store it at
4°C (unless you are aliquoting a newly arrived antibody; see next
section).
2. What to do with newly
arrived antibodies: ALIQUOT!
a. If the antibody arrives frozen, thaw it and
place on ice. If it arrives in liquid form, immediately place it on
ice. Vortex gently to mix, and spin briefly in microcentrifuge to
collect all liquid at bottom of tube.
b. Gently vortex the antibody solution to mix,
and spin briefly in microcentrifuge to collect all liquid at bottom
of tube.
c. Estimate the total volume of the antibody
solution, and how many aliquots you want to make. Typically,
aliquots of 10-50 ml are appropriate (larger aliquots for antibodies
that are used with relatively little dilution, e.g. 1:10; and
smaller aliquots for antibodies that will be highly diluted, e.g.
1:5000).
d.
Label
500 ml tubes with the antibody
name, the dilution, and the date aliquoted; e.g., anti–calretinin,
undiluted, 3/6/02.
e. Aliquot the antibody into the tubes. Put the
tubes in a box in the freezer, and make sure the box is
appropriately labeled (e.g., Hevner lab polyclonal antibodies).
f.
Enter the antibody in our computer database.
Put any accompanying literature or instructions in our antibody
notebook.
g. If you thaw an antibody, write the thaw date
on the tube; e.g., thaw 3/8/02.
3. Preparing antibodies
for immunohistochemistry
a.
Use thawed antibody stocks first,
but only if less than 3-6 months old.
b. If you need more antibody, thaw a new aliquot, write the thaw date on the tube, and thereafter store in the refrig. (i.e., do not re-freeze).