Journal of Lipid Research, Vol 38, 2603-2614, Copyright © 1997 by Lipid Research, Inc.
Protocol for the preparation of a segmental linear polyacrylamide gradient gel: simultaneous determination of Lp[a], LDL, and HDL particle sizes
X Li, W Innis-Whitehouse, WV Brown and NA Le
Department of Medicine, Emory University School of Medicine, Atlanta, GA 30322, USA.
We describe in this report a protocol for the preparation of a
polyacrylamide gel system (S-GGE 2.8/8.30) that consists of two linear
gradients designed for the simultaneous determination of the diameters of
LDL and HDL from whole plasma. The lower gel consists of an 8-30% linear
gradient which is optimum for the resolution of HDL subfractions and the
upper gel consists of a 2-8% linear gradient to allow for the resolution of
LDL and larger lipoprotein fractions such as Lp[a] and small VLDL. In
contrast to other non-denaturing gradient gel systems which are based on
protein staining, the present system uses lipid stain to specifically
identify lipoproteins. This approach also allows the plasma to be
pre-stained with immediate visualization of the lipid bands being possible
at the completion of the electrophoretic run. Using commercially available
gel casting equipment, the present gradient gel system can accommodate up
to 21 lanes per gel. The inter- run and intra-run coefficients of variation
for LDL particle size are 0.47 and 0.16%, respectively. The inter- and
intra-run CVs for Lp[a] particle size are 0.92% and 0.89%, respectively.
The inter-run and intra-run coefficients of variation for HDL2 and HDL3
particle size are 1.36% and 3.23%, respectively.
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