whole blood and amount of antibody(ies)

From: Maciej Simm (simmmmer@yahoo.com)
Date: Tue Oct 17 2000 - 20:36:22 EST


greetings everyone,

we have recently started using an anti-CD45 antibody which is very
potent and after titrating I determined 2.5 uL stock is optimal. I
noticed my coworker has trouble consistently distributing this small
volume across all the samples. I called the manifacturer about
possibly diluting the mAB and they recommended .1% NaN3. This
dilution (I am planning a ten-fold) would increase the final volume
of the staining mixture.

question - does increasing the volume of the staining mixture
(50-100uL whole blood, + 3 X 20uL of different mAB's) affect the
outcome of staining for each of the 3 antibodies present?

Any input, as always, is greately appreciated.

Maciej

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