Bio-Rad Profinia™ Protein Purification Instrument - Manual (Page 127)

Figure 10.1. shows proper placement of native IMAC buffers in the Profinia instrument.
Fig. 10.1. Native IMAC buffers properly positioned with instrument lid closed.
10.3 Denaturing IMAC Purification and Buffer Kits
The buffers provided in the denaturing IMAC kit are formulated with identical salts and
buffers as the native IMAC solutions, but require the addition of urea from the urea packs
provided with each kit. Denaturing IMAC buffers are used for proteins that partition into the
insoluble fraction of bacterial lysates or have inaccessible His sequence tags. The denaturing
IMAC buffers can also be used to solubilize the entire bacterial pellet without separation of
the soluble and insoluble fractions. Purifications from unfractionated bacterial pellets may
result in slightly less pure proteins. Table 10.3 provides a list of buffer compositions. The
lysis buffer is used for sample preparation (see Appendix E), and is not used in any of the
instrument ports. The IMAC cartridges are identical to the cartridges described in section
10.2, and all specifications apply to denaturing IMAC purifications.
120