Human ZAP70 knockout Jurkat cell line

• WB

Lab

Western blot - Human ZAP70 knockout Jurkat cell line (AB273841)

False colour image of Western blot : Anti-ZAP70 antibody [E267] staining at 1/500 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot ab32410 was shown to bind specifically to ZAP70. A band was observed at 70 kDa in wild-type Jurkat cell lysates with no signal observed at this size in ZAP70 CRISPR-Cas9 edited cell line ab273841 (CRISPR-Cas9 edited cell lysate ab273795). The band observed in the CRISPR-Cas9 edited lysate lane below 70 kDa is likely to represent a truncated form of ZAP70. 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 ZAP70 CRISPR-Cas9 edited Jurkat 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 °. 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® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-ZAP70 antibody [E267] (<a href='/products/primary-antibodies/zap70-antibody-e267-ab32410'>ab32410</a>) at 1/500 dilution

Lane 1:

Wild-type Jurkat cell lysate at 20 µg

Lane 2:

ZAP70 CRISPR-Cas9 edited Jurkat cell lysate at 20 µg

Lane 3:

MOLT-4 cell lysate at 20 µg

Lane 4:

Raji cell lysate at 20 µg

Predicted band size: 69 kDa

Observed band size: 70 kDa

false

• WB

Lab

Western blot - Human ZAP70 knockout Jurkat cell line (AB273841)

False colour image of Western blot : Anti-ZAP70 antibody [YE291] staining at 1/500 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot ab32429 was shown to bind specifically to ZAP70. A band was observed at 70 kDa in wild-type Jurkat cell lysates with no signal observed at this size in ZAP70 CRISPR-Cas9 edited cell line ab273841 (CRISPR-Cas9 edited cell lysate ab273795). The band observed in the CRISPR-Cas9 edited lysate lane below 70 kDa is likely to represent a truncated form of ZAP70. 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 ZAP70 CRISPR-Cas9 edited Jurkat 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 °. 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® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-ZAP70 antibody [YE291] (<a href='/products/primary-antibodies/zap70-antibody-ye291-ab32429'>ab32429</a>) at 1/500 dilution

Lane 1:

Wild-type Jurkat cell lysate at 20 µg

Lane 2:

ZAP70 CRISPR-Cas9 edited Jurkat cell lysate at 20 µg

Lane 2:

Western blot - Human ZAP70 knockout Jurkat cell line (ab273841)

Lane 3:

MOLT-4 cell lysate at 20 µg

Lane 4:

Raji cell lysate at 20 µg

Predicted band size: 69 kDa

Observed band size: 70 kDa

false

• NGS

Lab

Next Generation Sequencing - Human ZAP70 knockout Jurkat cell line (AB273841)

2 bp insertion after Pro90 of the WT protein