Determination of serum biomarkers of alcoholic fatty liver (AFL) using 2D-DIGE. (A) The experimental design used two individual samples each from healthy controls and AFL. (B) The combination of two different acquired images, in which, green color represents Cy3-labeled normal control and red color represents Cy5-labeled AFL. (C) The identified proteins are indicated by arrows on the 2D gel. Ultimately there were eight proteins selected according to statistical significance with a t-test value of < 0.05 (p < 0.05) as analyzed by the DeCyder software. Vitamin D-binding protein (VDBP), haptoglobin (HP), C-reactive protein (CRP), alpha-fetoprotein (AFP), inter-alpha-inhibitor H4 heavy chain (ITIH4), serine protease inhibitor Kazal-type 5 (SPINK5) and heak shock protein 75 kDa (HSP75) and afamin.