Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP with 15 min fixation) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Mouse IgG (Part No. 12-371B), or 2 µg of Anti-CTCF (Part No. CS204383) and the Magna ChIP™ G Kit (Cat. # 17-611).
Successful immunoprecipitation of CTCF associated DNA fragments was verified by qPCR using ChIP Primers, Human H19 ICR (Part No. CS204390) as a positive locus, and a representative published negative locus (Wendt, K.S., et al., 2008) (Figure 2). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

Western Blot Analysis:
Representative lot data.
K562 cell lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with Anti-CTCF, clone 1.2.1.5.4 (1 μg/mL).
Proteins were visualized using a Goat Anti-Mouse conjugated to HRP and a chemiluminescence detection system.
Arrow indicates CTCF (~130 kDa).

The following molecular weights have been reported in the literature: 180 kDa (CTCF-180), 130 kDa (CTCF-130), 82 kDa (CTCF-82), 70 kDa (CTCF-70). See Klenova, E.M., et al, (1997) Nucleic Acids Res. 25, 466–473 and Torrano,V., et al, (2006) Journal of Cell Science 119, 1746-1759) for additional details.
Please see molecular weight section for additional information.