Anti-NEK2 antibody [20/NEK2] - BSA and Azide free

• WB

Lab

Western blot - Anti-NEK2 antibody [20/NEK2] - BSA and Azide free (AB279722)

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 (IRDye^® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

All lanes:

Western blot - Anti-NEK2 antibody [20/NEK2] (<a href='/en-us/products/primary-antibodies/nek2-antibody-20-nek2-ab279717'>ab279717</a>) at 1/1000 dilution

Lanes 1 and 3:

Wild-type HeLa cell lysate at 20 µg

Lanes 2 and 4:

NEK2 knockout HeLa cell lysate at 20 µg

Lane 5:

K562 cell lysate at 20 µg

Lane 6:

Jurkat cell lysate at 20 µg

Lane 7:

MCF7 cell lysate at 20 µg

Predicted band size: 52 kDa

Observed band size: 51 kDa

false

• WB

Lab

Western blot - Anti-NEK2 antibody [20/NEK2] - BSA and Azide free (AB279722)

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.

All lanes:

Western blot - Anti-NEK2 antibody [20/NEK2] (<a href='/en-us/products/primary-antibodies/nek2-antibody-20-nek2-ab279717'>ab279717</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

NEK2 CRISPR-Cas9 edited HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human NEK2 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-nek2-knockout-hela-cell-line-ab273850'>ab273850</a>)

Lane 3:

K562 cell lysate at 20 µg

Lane 4:

Jurkat cell lysate at 20 µg

Predicted band size: 52 kDa

Observed band size: 50 kDa

false

• WB

Supplier Data

Western blot - Anti-NEK2 antibody [20/NEK2] - BSA and Azide free (AB279722)

This data was developed using ab279717, the same antibody clone in a different buffer formulation.

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

All lanes:

Western blot - Anti-NEK2 antibody [20/NEK2] (<a href='/en-us/products/primary-antibodies/nek2-antibody-20-nek2-ab279717'>ab279717</a>) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

Lane 2:

HFF-1 (human Skin fibroblast), whole cell lysate at 20 µg

Lane 3:

K-562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 52 kDa

Observed band size: 48 kDa

false