2nd step antibodies, simple beginner's error - maybe

From: Maciej Simm (simmmmer@yahoo.com)
Date: Fri Aug 04 2000 - 12:27:17 EST


Dear flowers,

well, I ran some more samples and here is what I found:

the monocytes sorted with magnetic anti-CD14 but NOT stained are a
tad brighter (2 clicks on the log scale) vs. the unstained
lymphocytes (sorry, no unstained monos in this batch) in both FL1 and
FL2 (I didn't study any other channels)

anti-isotype antibody  for the MACS anti-CD14 antibody pics up signal
in the 2nd decade (same instrument settings as isotype control on
monos)

this proves magnetic CD14 antibodies stay on the sorted cells and so
polyclonal secondary antibody should not be used when magnetic
separation is used (duh.)

I'll keep the group posted if anything else comes up along this
topic.

As far as recommended vendors of antibodies I asked for in the
original post, I'll compile a post next week b/c I'm still getting
new emails.

regards, Maciej



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