Bio-Rad Aurum Serum Protein Mini Kit - Manual (Page 6)

Section 5
Guidelines for Using the Aurum Serum
Protein Mini Kit
• Serum samples should be clarified before application to Aurum serum
protein columns.
• High salt concentrations (>200 mM) should be avoided due to
interference with albumin removal. Salt can also interfere with subsequent
IEF analysis. High salt samples can be dialyzed against a low-salt buffer
(25 mM phosphate, pH 7.0).
• For serum samples containing a high concentration of albumin, reduce
the sample load on the resin from 200 µl to 150 µl. The load may need to
be further optimized.
• For serum samples with low concentrations of albumin, increase the
sample load from 200 µl to 250 µl. An alternative would be to dilute the
initial sample 1/3 with binding buffer and apply 200–250 µl. The load may
need to be further optimized.
• The bound albumin and IgG can be recovered from the Aurum serum
protein columns and analyzed. For 1-D analysis, elute the column with
500 µl of Laemmli sample buffer (Bio-Rad catalog #161-0737). For 2-D
analysis, elute the column with 500 µl of ReadyPrep
TM sequential
extraction reagent 3 (Bio-Rad catalog #163-2104).
• If necessary the albumin/IgG-depleted samples can be concentrated
using a SpeedVac or lyophilizer.
• Albumin/IgG depleted serum samples, if not analyzed immediately, should
be stored at -20ºC.
• For 2-D electrophoresis analysis, see bulletin 2561, 2-D Electrophoresis
for Proteomics. A Methods and Product Manual.
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