Effect of different rehydration buffers on quality of western blotting analysis of 2D gels. 25 μg of mitotic chromosomes eluates were separated on pH6-11 IPG strips using sRB (A, B) or oRB (C, D) as in Figure 4. 2D gels were stained with SYPRO Ruby (A, C) or immunoblotted using anti-XCAP-E antibody (B, D). The protein spots seen on the top right of (A) were not reproducible and were perhaps due to poor alkylation. The spot pattern observed in (C) was reproducible.