Recombinant Anti-NEK2 antibody [20/NEK2] (ab279717)

False colour image of Western blot: Anti-NEK2 antibody [20/NEK2] staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab279717 was shown to bind specifically to NEK2. A band was observed at 51 kDa in wild-type HeLa cell lysates with no signal observed at this size in NEK2 CRISPR-Cas9 edited cell line ab266027 (NEK2 CRISPR-Cas9 edited cell lysate ab258070). The band observed in the CRISPR-Cas9 edited lysate lane above 51 kDa is likely to represent NEK2 with an insertion. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and NEK2 CRISPR-Cas9 edited HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1 % Tween^® 20 (TBS-T) 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-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDyeMCF7 cell lysateMCF7^® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

False colour image of Western blot: Anti-NEK2 antibody [20/NEK2] staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab279717 was shown to bind specifically to NEK2. A band was observed at 50 kDa in wild-type HeLa cell lysates with no signal observed at this size in NEK2 CRISPR-Cas9 edited cell line ab273850 (CRISPR-Cas9 edited cell lysate ab260448). The band observed in the CRISPR-Cas9 edited lysate lane below 50 kDa is likely to represent a truncated form of NEK2. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and NEK2 CRISPR-Cas9 edited HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) 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-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

Blocking and diluting buffer and concentration: 5% NFDM/TBST

The expression molecular weight observed is consistent with what has been described in the literature (PMID:25043295).

Negative control: HFF-1 (PMID:27509921).

Exposure time: 48 seconds