Figure 3 of
Khanobdee, Mol Vis 2004;
Figure 3. Detection of eukaryotically expressed proteins by the SH3BP4 antibody
A: Cell lysates from ARPE-19 (lanes 1 and 4), Y79 (lanes 2 and 5), COS-7 (lanes 3 and 6) and ARPE-19 transfected with GFP-SH3BP4 (lane 7) or myc-SH3BP4 (lane 8) constructs were analyzed by western blotting using a 1:10,000 dilution of either the chicken anti-human SH3BP4 IgY antibody (lanes 1-3, 7, and 8) or pre-immune IgY (lanes 4-6). Equal amounts of protein were loaded on each lane, and the amount of SH3BP4 was measured by the immunoreaction on the western blot. B: To quantify the level of 120 kDa SH3BP4, each blot was exposed to film and the resulting image scanned. The absorbance of each band was measured as an integrated optical density (IOD). The level of SH3BP4 protein from each cell line was quantitated in triplicate lysates as IOD/μg total protein. The height of the bars represent the mean of the fraction of the total SH3BP4 immunoreactivity from each set of lysates, averaged across the three blots. The standard deviation is illustrated by the error bars. The fold difference relative to the level in Y79 cells is indicated by the number above each bar. ARPE-19 levels were found to be significantly different from Y79 levels (p<0.001) and COS-7 levels (p<0.001) using analysis of variance with the Student-Newman-Keuls post hoc test.