Anti-CD3 zeta antibody

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD3 zeta antibody (AB190728)

Immunofluorescence analysis of A549 cells, using CD3 Zeta polyclonal antibody at 1 : 50.

• WB

Lab

Western blot - Anti-CD3 zeta antibody (AB190728)

False colour image of Western blot : Anti-CD3 zeta antibody staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab190728 was shown to bind specifically to CD3 zeta. A band was observed at 16 kDa in wild-type Jurkat cell lysates with no signal observed at this size in CD247 knockout cell line ab273856 (knockout cell lysate ab273810). To generate this image, wild-type and CD247 knockout 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°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^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-CD3 zeta antibody (ab190728) at 1/500 dilution

Lane 1:

Wild-type Jurkat cell lysate at 20 µg

Lane 2:

CD247 knockout Jurkat cell lysate at 20 µg

Lane 2:

Western blot - Human CD247 knockout Jurkat cell line (<a href='/en-us/products/cell-lines/human-cd247-knockout-jurkat-cell-line-ab273856'>ab273856</a>)

Lane 3:

MOLT-4 cell lysate at 20 µg

Lane 4:

K562 cell lysate at 20 µg

Predicted band size: 19 kDa

Observed band size: 16 kDa

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• WB

Supplier Data

Western blot - Anti-CD3 zeta antibody (AB190728)

All lanes:

Western blot - Anti-CD3 zeta antibody (ab190728)

All lanes:

extracts of Jurkat cell line

Predicted band size: 19 kDa

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