Fig_2.tif (4.23 MB)

Example of two dimensional gel electrophoresis (2D-GE) analyses comparing control vs. AMD sera.

figure

posted on 2016-01-05, 14:53 authored by Alessandro Iannaccone, Francesco Giorgianni, David D. New, T. J. Hollingsworth, Allison Umfress, Albert H. Alhatem, Indira Neeli, Nataliya I. Lenchik, Barbara J. Jennings, Jorge I. Calzada, Suzanne Satterfield, Dennis Mathews, Rocio I. Diaz, Tamara Harris, Karen C. Johnson, Steve Charles, Stephen B. Kritchevsky, Ivan C. Gerling, Sarka Beranova-Giorgianni, Marko Z. Radic

The left panel shows a gel on which protein from human macular lysate immunoprecipitated by a control serum sample have been separated on two dimensions (by molecular weight and pI), compared to an AMD sample ran identically and simultaneously shown on the right. There are several spots on the 2D-GE AMD gel that appear different compared to the control one. Confirmed antigens recognized by the AAbs present in the serum of this one particular AMD participant included HSPA8, ANXA5 and HSPB4/CRYAA (arrows). See text for further discussion of these findings. Note also how the AMD gel exhibits a stronger “load” compared to the control gel, consistent with the fact that AMD sera tended to be more often autoreactive and thus, even when some autoreactivities were shared, AMD sera exhibited much stronger levels of reactivity, indicative of a larger amount and numbers of proteins pulled down by the IP process.