Dear All,
I am asking for the most valid way of comparing different cell lines for
the expression of a membrane bound protein (MRP1). The expression of this
protein is very low generally but we have not as yet found a truly negative
cell line.
The isotype control is used for each of the samples. We use one cell line
(K562) as a standard and calculate ratios of expression relative to this.
However, the isotype control varies for each of the cell lines.
My question is - do we set the K562 isotype control at a level that allows
all isotype controls to be measurable in the mid or above first decade of a
log scale or do we set it somewhere close to the first decade?
Below is a table of MFI showing what happens for the same samples for each
scenario.
Iso control MRP1 Iso control MRP1
K562 13.23 32.89 33.49 71.03
HL60 5.01 15.09 13.07 36.55
CEM 3.23 12.78 7.49 33.25
MOLT4 1.93 9.91 4.96 25.03
U266 3.62 13.27 9.68 32.80
We calculate the MRP1 value by MRP1(MFI) - iso control (MFI). The ratios
are not equivalent for the 2 data sets. Any advice or agreed dogma on this
issue would be appreciated.
Many thanks, Bryone
Dr. Bryone J Kuss
MB BS PhD (Adelaide)
FRACP FRCPA (Haematology)
Peter Nelson Leukaemia Research Fellow
Dept. of Haematology and Genetic Pathology
Flinders Medical Centre
Bedford Park
South Australia 5042
ph. 618 8204 3981 (office) 618 8204 4392 (laboratory)
fax. 618 8204 5450
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