Rat CD45, a correction and some zinc formalin/Zinc TRIS buffer fixative, again!
|From:||Gayle Callis <email@example.com>|
The rat CD45R (Pharmingen) was done on acetone fixed frozen sections and
not FFPE sections. FFPE can work on murine CD45R with enzyme digestion. My
file didn't have anything about CD45 (CLA).
Now for a discourse on zinc containing fixatives: You may have seen this
before, so delete it now or risk boredom.
Pharmingen has left one thing very unclear in its description of "zinc
fixation" when describing this fixation needed for an antibody. Confusion
may begin here!
When BD-Pharm refers to zinc fixation, this is NOT zinc formalin. This is
a formalin free fixative developed by J. Beckstead for human lymphoma cell
marker studies/paraffin sections, and then by Nitta et al for murine cell
surface markers. IT is a zinc TRIS buffer which preserves the morphology
adequately enough for paraffin work, and alk phosphatase method must be
used with it, since endog peroxidase CAN"T be quenched. No antigen
retrieval is needed and antibody concentrations similar to frozen section
IHC can be used.
It is my dearest wish that someday, somehow - Pharmingen will clear up the
naming of this "zinc fixative" so it does not get mixed up with zinc
formalin, might help people out before they stuff a tissue into zinc
formalin by mistake and screw up an experiment. It happens, folks!
Call it Zinc TRIS buffer fixative or whatever. PLEASE HELPPPPPPP!!!! are
they listening in???? We are so attuned to zinc formalin, it is too easy
to misread something that says, very generally, zinc fixation. CHANGE IT!
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
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