Bio-Rad Chromatographic Surfaces - Manual (Page 2)

Uses
I Protein profiling and biomarker discovery
I Rapid protein analysis to determine purity, mass confirmation,
or both
How It Works
The ProteinChip H50 array surface binds proteins through reverse-
phase or hydrophobic interaction chromatography and has binding
characteristics similar to that of a C6 to C12 alkyl chromatographic
resin. In reverse-phase interactions, proteins within the sample are
partitioned between the lipophilic phase of the array surface and
the sample buffer. Proteins less hydrophobic relative to the binding
buffer will not bind to the array surface, while proteins more
hydrophobic will bind to the array surface.
By increasing the organic content of the washing buffer, the
hydrophobic nature of the buffer increases. Proteins that had
previously bound to the array will repartition into the washing buffer
and be washed away if their hydrophobicity is less than that of the
washing buffer. Only the most hydrophobic proteins will be retained
with wash buffers containing a high organic solvent.
Hydrophobic interaction chromatography is characterized by
binding of proteins to a hydrophobic surface at high salt
concentrations (salt precipitation of proteins). Typically conditions
are nondenaturing, and since no organic solvent is used, biological
activity has a much higher probability of being retained. Proteins are
sequentially washed from the array surface by decreasing the salt
concentration of the wash buffers.
Packaging and Storage
Store the arrays at room temperature.
ProteinChip arrays are packaged in a 12-array cassette.
A bioprocessor reservoir is included in the package (see Figure 1).
The spare ProteinChip cassette included to separate the reservoir
from the arrays should be removed before use in the ProteinChip
cassette-compatible bioprocessor (catalog #C50-30011). It is not
necessary to remove the arrays when using the cassette-compatible
© 2006 Bio-Rad Laboratories, Inc.