Why LS column from Miltenyi eat away my cells

chen li chen_li3 at yahoo.com
Tue Jun 26 08:08:37 EDT 2007


Dear Flowers,

Many of you might also use microbeads and columns from
Miltenyi to preform either positive and negative
selection. Yesterday I did a negative selection using
LS column. After staining the cells from 5 day one-way
mixed lymphocyte reaction with biotin-conjugated
anti-human mAb, out of curiosity, I passed the cells
without incubating them with anti-biotin
microbeads.The cells number after staining but before
loading onto LS column is 30x10e6, and the cells in
the flow-through(after going through column) is
20x10e6 cells. It looks like the column EAT AWAy/trap
many cells even without adding microbeads.  Even worse
I got only 0.5x10e6 cells after adding anti-biotin
microbeads (the expected negative fraction is about
5-10x10e6 cells). I  wonder if anyone notice that a
lot of cells are lost after passing through the column
and how you prevent cell loss. 


Thank you very much,

Li


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