Bio-Rad Bio-Gel P Polyacrylamide Gel - Manual (Page 8)

After placement of the buffer bottles and the cartridge lines have been
automatically primed, the cartridges are ready to insert into the instrument.
They are automatically equilibrated by the buffers while running a
preprogrammed method. Remove the luer caps and screw cartridges into
the cartridge holders on the instrument. Be careful not to overtighten
cartridges. Refer to Figure 2 for the proper connection of the cartridges in
the Profinia system. The female luer inlet (top) and the male luer outlet
(bottom) simplify the orientation and connection. At the end of the run, the
cartridges are equilibrated into storage solution, and should be recapped
with the luer fittings. After use, the cartridges should be stored at 4°C and
be reused within 6 months.
1. 2.
3. 4.
Fig. 2. Bio-Scale™ Mini cartridge installation procedure.
Section 3
Denaturing IMAC Purification
3.1 General Buffer and Cartridge Information
The buffers used in the denaturing IMAC purification are identical to
the native IMAC solutions, but require the addition of urea. We do not
provide ready-to-use denaturing IMAC purification kits. Denaturing IMAC
buffers are used for proteins that partition into the insoluble fraction of
E. coli lysates or have inaccessible His sequence tags. The denaturing
IMAC buffers can also be used to solubilize the entire
E. coli pellet without
separation of the soluble and insoluble fractions. Purifications from
unfractionated
E. coli pellets may result in slightly less pure proteins.
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1. Prior to runs, the cartridge holders are
maintained in the connected position
for storage.
2. Unscrew and separate column
connection holders so that they are
ready to receive purification cartridges.
Remove the bottom luer cap of an
affinity cartridge and insert it into
cartridge position #1 (left).
3. Pull the top portion of the column
holder down and screw closed to
create a seal for cartridge #1. Be
careful not to overtighten cartridges.
4. Desalting applications for native IMAC
and GST require the addition of the
desalting cartridge in cartridge position
#2 (right).
1 2 1 2
1 2 1 2
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