Anti-Nucleolin antibody [EPR7951] - BSA and Azide free

• WB

Lab

Western blot - Anti-Nucleolin antibody [EPR7951] - BSA and Azide free (AB248709)

This data was developed using ab134164, the same antibody clone in a different buffer formulation.

Lanes 1 - 3 : Merged signal (red and green). Green - ab134164 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab134164 was shown to specifically react with Nucleolin in wild-type HeLa cells as signal was lost in Nucleolin knockout cells. Wild-type and Nucleolin knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab134164 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Nucleolin antibody [EPR7951] (<a href='/en-us/products/primary-antibodies/nucleolin-antibody-epr7951-ab134164'>ab134164</a>) at 1/10000 dilution

Lane 1:

Wild-type HeLa whole cell lysate at 20 µg

Lane 2:

Nucleolin knockout HeLa whole cell lysate at 20 µg

Lane 3:

MCF7 whole cell lysate at 20 µg

Predicted band size: 76 kDa

Observed band size: 100 kDa

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• WB

Unknown

Western blot - Anti-Nucleolin antibody [EPR7951] - BSA and Azide free (AB248709)

This data was developed using ab134164, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Nucleolin antibody [EPR7951] (<a href='/en-us/products/primary-antibodies/nucleolin-antibody-epr7951-ab134164'>ab134164</a>) at 1/1000 dilution

Lane 1:

K562 cell lysates at 10 µg

Lane 2:

HuT-78 cell lysates at 10 µg

Lane 3:

HeLa cell lysates at 10 µg

Lane 4:

Jurkat cell lysates at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 76 kDa

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• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-Nucleolin antibody [EPR7951] - BSA and Azide free (AB248709)

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.