All lanes : Anti-CTCF antibody [mAbcam 37477] (ab37477) at 5 µg/ml

Lane 1 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 83 kDa


Exposure time: 90 seconds

All lanes : Anti-CTCF antibody [mAbcam 37477] (ab37477) at 2 µg/ml

Lane 1 : ZR-75-1 cell extract
Lane 2 : HCT-116 cell extract

Predicted band size: 83 kDa



ab37477 recognises a band at approximately 130 kDa corresponding to CTCF.

Overlay histogram showing HeLa cells stained with ab37477 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab37477, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.