re: freezing antibodies

From:Jan Shivers

My method of freezing and storing Abs:

First I look to see if the antibody data sheet from the manufacturer states
that the antibody HAS to be stored frozen.  If that is the case, then I work
up the antibody (while it's in the unfrozen state at arrival), to determine
the optimal antibody dilution, et al.  Once I know the optimal dilution, I
know what volume of an antibody aliquot I'd mostly likely need to mix up a
fresh working dilution in the future.

For instance, if I work up an antibody and find that a dilution of 1:50
works best, then I know that whenever I mix up 1 ml of working dilution I
will need 20 ul of antibody.... so..... I then put aliquots of 25 ul each
into microcentrifuge tubes and store them in a Sarstedt cardboard box in the
freezer.  I always pipet 5 more uls into each microcentrifuge tube than I
will need in the future, because when I go to retrieve (pipet) the antibody
out of the tube at a later date, I can never pipet out every single ul.
Thus, the reason for pipeting in a little more than I need.

The few ul remainder is put back in the freezer box with a red dot on the
top of the vial, telling me that it has been thawed once already.  These are
held as emergency back-ups, should I ever run out of Ab before my next
shipment comes in.

This may not be terribly cost-efficient, but it is easier for me to waste a
few uls here and there than to add buffer to the vial and attempt to flush
out all the antibody that has clung to the side of the vial.

Jan Shivers
U of MN Vet Diag Lab





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