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All lanes : Anti-FCP1 antibody (ab52722) at 1/1000 dilution
Lane 1 : 20µg of a total protein extract from E coli with ~50ng to 500ng of a tagged fusion protein of an irrelevant antigen.
Lane 2 : 20µg of a total protein extract from E coli with ~50ng to 500ng of the antigen (Tagged fusion protein).
Secondary
Rabbit anti-mouse IgG + IgM, (H+L) HRP conjugated, at 1/5000 dilution
Predicted band size : 104 kDa
Observed band size : 38 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 enzyme tag.
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