Human CRY1 (Cryptochrome I) knockout HeLa cell lysate

• WB

Lab

Western blot - Human CRY1 (Cryptochrome I) knockout HeLa cell lysate (AB258382)

Lane 1 : Wild-type HeLa cell lysate 20 ug
Lane 2 : CRY1 knockout HeLa cell lysate 20 ug
Lane 3 : HCT116 cell lysate 20 ug
Lane 4 : Caco-2 cell lysate 20 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab245564 observed at 66 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab245564 was shown to react with Cryptochrome I/CRY1 in wild-type HeLa cells in Western blot with loss of signal observed in CRY1 knockout cell line ab265791 (CRY1 knockout cell lysate ab258382). Wild-type HeLa and CRY1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab245564 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 ^°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-Cryptochrome I/CRY1 antibody (<a href='/en-us/products/primary-antibodies/cryptochrome-i-cry1-antibody-ab245564'>ab245564</a>) at 1/2000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CRY1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CRY1 (Cryptochrome I) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-cry1-cryptochrome-i-knockout-hela-cell-line-ab265791'>ab265791</a>)

Lane 3:

HCT116 cell lysate at 20 µg

Lane 4:

Caco-2 cell lysate at 20 µg

Predicted band size: 66 kDa

Observed band size: 66 kDa

false

• Sanger seq

Unknown

Sanger Sequencing - Human CRY1 (Cryptochrome I) knockout HeLa cell lysate (AB258382)

Homozygous : 13 bp deletion in exon1