Abstract
Purpose: :
To determine the presence of alpha crystallin inside lipofuscin from human retinal pigment epithelium.
Methods: :
1) Human lipofuscin granules were purified from RPE cells by sedimentation and ultracentrifugation in a discontinuous sucrose gradient, by using three times the protocol described previously and adding one step of "small discontinuous gradient". This method was used to obtain a less dirty lipofuscin from cellular debries. 2) Human alpha crystallin was identified by western blotting, mass spectrometry and immunogold electron microscopy.
Results: :
The three techniques were useful to find out the alpha crystallin protein in the lipofuscin granules.
Conclusions: :
Alpha crystalln is present in the lipofuscin granules from human RPE.
Keywords: crystallins • retinal pigment epithelium • ipofuscin