Bio-Rad Foresight™ Chromatography Columns, Prepacked - Manual (Page 30)

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3.6.4 Washing Weakly Bound Material
• Add 300 μl of wash buffer per well and
cover the plate with sealing film. Incubate
for 1 min at 1100 rpm. Next, transfer
the media and collection plate to the
centrifuge for liquid removal. Run a cycle
at 300 g for typically 1–2 min. Do not
use vacuum filtration for this step. After
the liquid has been collected, label the
collection plate “Wash”, blot the media
plate, and place it on top of a new
collection plate
• The Wash collection plate can now be
analyzed. Additional washes can be
performed; each should be collected in
its own collection plate
3.6.5 Elution
• Add 300 μl of elution buffer per well
and cover the plate with sealing film.
Incubate for 1 min at 1100 rpm. Next,
transfer the media and collection plate
to the centrifuge for liquid removal. Run
a cycle at 300 g for at least 2 min. Do
not use vacuum filtration for this step.
After the liquid has been collected, label
the collection plate “Elution_1”, blot the
media plate, and place it on top of a new
collection plate (Elution_2)
• Usually three elution cycles are sufficient
to recover the target molecule. Each
elution cycle requires its own collection
plate