Human CDKN1C (p57 Kip2) knockout HeLa cell lysate

• WB

Lab

Western blot - Human CDKN1C (p57 Kip2) knockout HeLa cell lysate (AB280120)

Lane 1 : Wild-type HeLa Vehicle Control Dexamethasone (0 nM, 16 h) ab277359 cell lysate 20 μg
Lane 2 : Wild-type HeLa Treated Dexamethasone (50 nM, 16 h) ab287335 cell lysate 20 μg
Lane 3 : CDKN1C knockout HeLa Vehicle Control Dexamethasone (0 nM, 16 h) ab277299 cell lysate 20 μg
Lane 4 : CDKN1C knockout HeLa Treated Dexamethasone (50 nM, 16 h) ab281877 cell lysate 20 μg
Lane 5 : MCF7 cell lysate 20 μg
Lane 6 : SH-SY5Y cell lysate 20 μg
False colour image of Western blot : Anti-p57 Kip2 antibody [EP2516] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab119989 was shown to bind specifically to p57 Kip2. A band was observed at 50 kDa in wild-type HeLa cell lysates with no signal observed at this size in CDKN1C knockout cell line ab280061 (knockout cell lysate ab280120). To generate this image, wild-type and CDKN1C knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) at 1/20000 dilution.

All lanes:

Western blot - Anti-p57 Kip2 antibody [EP2516] (<a href='/en-us/products/primary-antibodies/p57-kip2-antibody-ep2516-ab119989'>ab119989</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa Vehicle Control Dexamethasone (0 nM, 16 h) ab277359 cell lysate at 20 µg

Lane 2:

Wild-type HeLa Treated Dexamethasone (50 nM, 16 h) ab287335 cell lysate at 20 µg

Lane 2:

Western blot - Human CDKN1C (p57 Kip2) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-cdkn1c-p57-kip2-knockout-hela-cell-line-ab280061'>ab280061</a>)

Lane 3:

CDKN1C knockout HeLa Vehicle Control Dexamethasone (0 nM, 16 h) ab277299 cell lysate at 20 µg

Lane 4:

CDKN1C knockout HeLa Treated Dexamethasone (50 nM, 16 h) ab281877 cell lysate at 20 µg

Lane 5:

MCF7 cell lysate at 20 µg

Lane 6:

SH-SY5Y cell lysate at 20 µg

Predicted band size: 32 kDa

Observed band size: 50 kDa

false

• Sanger seq

Lab

Sanger Sequencing - Human CDKN1C (p57 Kip2) knockout HeLa cell lysate (AB280120)

217 bp Deletion in Exon 1