Anti-p29 ING4 polyclonal antibody detects ING4 protein by western blot. This antibody was used at 1.0 ug/ml to detect ING4 (lane 2) present in a U2OS whole cell lysate over expressing the protein. A control lysate (lane 3) shows no background staining. Comparison to MW markers (lane 1) indicates detection of a single band at ~29 kDa corresponding to ING4.

A 4-20% Tris-glycine gradient gel was used to separate the protein by SDS-PAGE under reducing conditions. The protein was transferred to nitrocellulose using standard methods. After blocking using 5% non-fat dry milk in PBS, the membrane was probed with the primary antibody overnight at 4° C followed by washes and reaction with a 1:20,000 dilution of IRDye800 conjugated Rabbit anti Goat IgG [H&L] for 45 min at room temperature. LICOR's Odyssey® Infrared Imaging System was used to scan and process the image. Other detection systems will yield similar results.

Anti-p29 ING4 polyclonal antibody detects ING4 protein by western blot in over expressed cell lysates. This antibody was used at 1.0 µg/ml to detect ING4 expression in control (-) and transformed U2OS and HeLa cell lysates. A predominant band corresponding to p29 ING4 is only seen in lysates from transformed cells. Anti-p29 ING4 polyclonal antibody detects ING4 protein by western blot in over expressed cell lysates. This antibody was used at 1.0 ug/ml to detect ING4 expression in control (-) and transformed U2OS and HeLa cell lysates. A predominant band corresponding to p29 ING4 is only seen in lysates from transformed cells.