Bio-Rad Profinia Protein Purification System Kits - Manual (Page 17)

6.2 Analysis of Purified Protein
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The standard report also summarizes
key run parameters such as time to
elution, flow rate, wash time, and
volume loaded. If the software
package is not part of your Profinia
system, the end of run summary
screen presents the key sample data
directly on the user interface screen
(Figure 14).
Fig. 14. Data report screen on user interface
screen.
Purified affinity-tagged proteins are
typically analyzed by gel
electrophoresis. For IMAC
purifications, samples of starting
material (L, resuspended lysate, or
load), flowthrough (FT, tube A),
wash 1 (W-1, tube B), wash 2 (W-2,
tube C), and purified protein (E, tube
D) are diluted 1:7 in Laemmli buffer,
boiled, and 10–20 µl is then loaded
onto a gel. The IMAC buffers contain
potassium salts and must be diluted
at least 1:7 to prevent potassium-SDS
precipitates from forming in Laemmli
buffer. The gel in Figure15 shows a
representative purification from the
51-D IMAC purification.
Fig. 15. Gel analysis of purified 51-D with
native IMAC.
L FT W-1 W-2 E
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