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All lanes : Anti- KAP1 antibody (ab43760) at 1/1000 dilution
Lane 1 : Total protein extract from E coli with ~50ng to 100ng of GST fusion protein of an irrelevant antigen at 20 µg
Lane 2 : Total protein extract from E coli with ~50ng to 500ng of the antigen (GST-antigen fusion protein)
Secondary
Rabbit anti-mouse IgG + IgM, (H+L) conjugated to Horseradish peroxidase at 1/5000 dilution
Predicted band size : 89 kDa
Observed band size : 38 kDa (why is the actual band size different from the predicted?)
Note: 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.
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