重组Anti-CSK抗体[EPR6771] (ab125005)

Western blot: Anti-CSK antibody [EPR6771] (ab125005) staining at 1/5000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab125005 was shown to bind specifically to CSK. A band was observed at 50 kDa in wild-type A549 cell lysates with no signal observed at this size in CSK knockout cell line. To generate this image, wild-type and CSK knockout A549 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: CSK knockout HAP1 cell lysate (20 µg)
Lane 3: Ramos cell lysate (20 µg)
Lane 4: Jurkat cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab125005 observed at 50 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab125005 was shown to specifically react with CSK when CSK knockout samples were used. Wild-type and CSK knockout samples were subjected to SDS-PAGE. ab125005 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

ab125005 at 1/50 dilution staining CSK in paraffin-embedded Human tonsil tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.