Lane 1: Wild-type HeLa cell lysate (20µg)

Lane 2: TRIM24 knockout HeLa cell lysate (20µg)

Lane 3: Hap1 cell lysate (20µg)

Lane 4: A549 cell lysate (20µg)

Lanes 1- 4: Merged signal (red and green). Green - ab256491 observed at 140 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab256491 Anti-TRIM24 antibody [EPR22825-2] was shown to specifically react with Tripartite Motif Containing 24 in wild-type HeLa cells in western blot. The band observed in the knockout cell line ab264963 (knockout cell lysate ab258246) lane below 140kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and Tripartite Motif Containing 24 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab256491 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4^°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

Allele-1: 2 bp insertion in exon 1

Allele-2: Insertion of the selection cassette in exon 1