All lanes : Anti-TR4 antibody [EPR1773(2)] (ab109301) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : NR2C2 knockout HEK293T cell lysate
Lane 3 : HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 65 kDa
Observed band size: 67 kDa why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab109301 observed at 67 kDa. Red - loading control ab8245 observed at 36 kDa.

 ab109301 Anti-TR4 antibody [EPR1773(2)] was shown to specifically react with TR4 in wild-type HEK293T cells. The band observed in knockout cell line ab266228 (knockout cell lysate ab257563) lane below 67 kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and TR4 knockout samples were subjected to SDS-PAGE. ab109301 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution 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: 16 bp deletion in exon6

 

Allele-2: Insertion of the selection cassette in exon 6.