Bio-Rad ProteoMiner Protein Enrichment Kits - Manual (Page 6)

Section 4
Reagent Preparation
Prepare elution reagent by adding 610 µl rehydration reagent to one vial
lyophilized elution reagent. Following rehydration, each vial will contain enough
elution reagent for processing two samples. If preparing an uneven number of
columns, you will have remaining material that will be required for subsequent
preparations. Remaining material may be stored at –20°C for up to one week.
However, it is recommended to make this solution fresh each time you use the
kit.
Section 5
Sample Considerations
This kit has been optimized for plasma (nonheparinized) and serum samples.
Sample loading and buffer compatibility have not been validated for other
sample types. However, ProteoMiner™ technology has successfully been
applied to other sample types including urine (Righetti et al. 2005), bile
(Housset et al. 2007), platelets (Boschetti et al. 2008), red blood cell extract
(Boschetti et al. 2008), and egg white extract (Boschetti et al. 2008).
Results with other sample types will vary depending on the amount of protein
in your sample; best results are obtained with 50 mg protein for the large-capacity
kits and 10 mg for the small-capacity kits. Caution: When working with
human plasma/serum it is important to follow biohazardous
material handling guidelines.
The ratio of protein to beads is crucial for optimal performance of ProteoMiner
kits. The dynamic range of the protein concentration in the sample is reduced
when the high-abundant proteins saturate their ligands and the low-abundant
proteins bind to a sufficient number of ligands to allow for enrichment.
Therefore, in order to achieve optimal results, it is important to load the
recommended amount of protein.
Section 6
Instructions for Use With ProteoMiner™
Large-Capacity Kits (Catalog #s 163-3007
and 163-3009)
This protocol has been optimized for plasma and serum samples with
protein concentrations of >50 mg/ml (requires total protein load >50 mg). For
other sample types, please refer to Section 5: Sample
Considerations.
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