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My name is David and I supervise the operation of a Coulter Elite. We've
done quite a bit with it including DNA analysis and sorting and would like
to know of other "Elite" operators throughout for potential exchange of
ideas and operating tips.
If I may, one simple question, we have (relatively speaking) little trouble
in labeling human cells with the variety of mouse anti human reagents,
either single step labeling or double (mouse MAb as primary followed by
GAM-FITC as secondary). Normally, our second antibody alone control
histograms are superimposable with those of an irrelevant primary ---
EXCEPT when we try to use rabbit-anti-human any marker you want serum.
Then all proverbial hell breaks loose and non-immune rabbit serum has a
histogram that almost matches our immune anti-serum. Our antisera were
raised in rabbits against a KLH-coupled peptide and does show specificity
against peptide alone. Western blots (for one of the three rabbit sera)
have demonstrated that our sera recognizes what it is supposed to
recongize. So it seems as though we may (?) have a lot of non-specific
attachment of rabbit antisera to our human cells... Can anyone help?
Fc-receptors?
I claim to be a card-carrying immunologist but this little problem has got
me stumped as well as 4 other otherwise educated folks in our lab. Is
there something simple we're missing? Something dumb we overlooked?
Second question: if there is a FAQ as there is for some of the bionet
groups, would some kind soul tell me it's location?
Many thanks in advance,
Yours,
David
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+ David L. Haviland, Ph.D. Internet:"haviland@kids.wustl.edu" +
+ Washington Univ. School of Med. A.K.A : The Compiler +
+ Dept. of Peds./Pulm. Box 8116 ICBM-Net : Just hit St. Louis +
+ 400 S. Kingshighway &-6 <- User is Brain Dead... +
+ St. Louis, MO 63110 FAX: 314-454-2476 +
+ (314) 454-6076 +
+ ------------------------------------------------------------------- +
+ REALITY.SYS - File Corrupted! Re-BOOT Universe? (Y/N)... +
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