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All lanes : Anti-PAK2 antibody (ab52811) at 1/1000 dilution
Lane 1 : ~20ug of a total protein extract from E coli with ~50ng to 100ng of a tagged fusion protein of an irrelevant antigen.
Lane 2 : ~20ug of a total protein extract from E coli with ~50ng to 500ng of the antigen (tagged-PAK2 fusion protein).
Secondary
HRP conjugated Rabbit anti-mouse IgG + IgM (H+L) at 1/5000 dilution
Predicted band size : 58 kDa
Observed band size : 32.5 kDa (why is the actual band size different from the predicted?)
The molecular weight of the band on the western blot does not correspond to the molecular weight of the natural protein because only a fragment of the gene is used and it is fused to an enzyme tag.
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