Bio-Rad ProteinChip Serum Fractionation Kits - Manual (Page 2)

© 2006 Bio-Rad Laboratories, Inc. © 2006 Bio-Rad Laboratories, Inc.
Introduction
The ProteinChip serum fractionation kit is designed to facilitate the
analysis of crude serum samples by fractionating proteins based on
their biophysical properties.
The kit allows high-throughput fractionation by using anion exchange
media in a 96-well microplate format. The anion exchange support is
supplied in a 96-well ProteinChip Q filtration plate and requires
rehydration before use. The samples are added to the plate and then
eluted in a stepwise manner by altering the pH of the wash buffer
until six fractions are collected. The fractions can then be analyzed
on a ProteinChip array using a profiling protocol. Each of the six
fractions is collected twice, and the two collections are pooled. This
helps to ensure that the pH changes appropriately, and also results in
greater reproducibility in the fractionation.
The fractions can be analyzed in your particular profiling application.
If you are using multiple array types or conditions, we recommend
that the same fraction is profiled at one time to avoid multiple
freeze-thaw cycles.
Materials
Materials Included
I ProteinChip U9 buffer (9 M urea, 2% CHAPS, 50 mM Tris-HCl,
pH 9), 20 ml
I Rehydration buffer (50 mM Tris-HCl, pH 9), 250 ml
I Wash buffer 1 (50 mM Tris-HCl, 0.1% OGP, pH 9), 20 ml
I Wash buffer 2 (50 mM HEPES, 0.1% OGP, pH 7), 30 ml
I Wash buffer 3 (100 mM Na acetate, 0.1% OGP, pH 5), 30 ml
I Wash buffer 4 (100 mM Na acetate, 0.1% OGP, pH 4), 30 ml
I Wash buffer 5 (50 mM Na citrate, 0.1% OGP, pH 3), 30 ml
I Wash buffer 6 (33.3% isopropanol, 16.7% acetonitrile,
0.1% trifluoroacetic acid), 30 ml
I ProteinChip Q filtration plate, filled with dehydrated anion
exchange Q ceramic HyperD F sorbent, 1