Anti-CSK antibody [EPR6771]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CSK antibody [EPR6771] (AB125005)

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.

• WB

Lab

Western blot - Anti-CSK antibody [EPR6771] (AB125005)

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.

All lanes:

Western blot - Anti-CSK antibody [EPR6771] (ab125005)

Predicted band size: 50 kDa

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• WB

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Western blot - Anti-CSK antibody [EPR6771] (AB125005)

All lanes:

Western blot - Anti-CSK antibody [EPR6771] (ab125005) at 1/1000 dilution

Lane 1:

Jurkat cell lysate at 10 µg

Lane 2:

Ramos cell lysate at 10 µg

Lane 3:

RAW 264.7 cell lysate at 10 µg

Secondary

All lanes:

Goat anti-Rabbit HRP at 1/2000 dilution

Predicted band size: 50 kDa

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• WB

Lab

Western blot - Anti-CSK antibody [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.

All lanes:

Western blot - Anti-CSK antibody [EPR6771] (ab125005) at 1/5000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

CSK knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human CSK knockout A549 cell line (<a href='/en-us/products/cell-lines/human-csk-knockout-a549-cell-line-ab300902'>ab300902</a>)

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 50 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-CSK antibody [EPR6771] (AB125005)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.