Harry Potter cloned a human gene (Muggle1) into a bacterial expansion vector so

Question

Harry Potter cloned a human gene (Muggle1) into a bacterial expansion vector so that there is a myc tag. an eterokinase site, and a histidine tag at the C-Terminus (a myc tag is a small amino acid sequence that can illicit an immune response). After inducing expression and applying the crude vacterial extract over a NI+2-agrose column, he subjected the effluent to both SDS-PAGE/Coomasi Blue and Western Blot analysis. The SDS-PAGE gel shows a prominent band around 70kD with several intese bands at 45kD, 33kD, and 25kD.

What are the ebst antibodies to detect the recomibant Muggle1 protein while at the same time determine if the lower bands are products of degradtion at the N-Terminus as opposed to the co-purfiying contaminatens

Mark your selection in the table below.

If more than one antibody is used indicate the order of use (1,2,3, ect...)

Goat anti myc tag Fish anti-histidne tag Donkey anti-histidine tag Eterokinase anti-chicken IgG Donkey anti-myc tag Harry Potter cloned a human gene (Muggle1) into a bacterial expansion vector so that there is a myc tag. an eterokinase site, and a histidine tag at the C-Terminus (a myc tag is a small amino acid sequence that can illicit an immune response). After inducing expression and applying the crude vacterial extract over a NI+2-agrose column, he subjected the effluent to both SDS-PAGE/Coomasi Blue and Western Blot analysis. The SDS-PAGE gel shows a prominent band around 70kD with several intese bands at 45kD, 33kD, and 25kD. What are the ebst antibodies to detect the recomibant Muggle1 protein while at the same time determine if the lower bands are products of degradtion at the N-Terminus as opposed to the co-purfiying contaminatens Mark your selection in the table below. If more than one antibody is used indicate the order of use (1,2,3, ect...)

Explanation / Answer

1) one antibody to bind to myc tag i.e. goat anti myc tag antibody

and 2)one to bind to muggle 1 protein region. fish anti muggle 1 antibody.

thus, we can find out if the smaller fragments in lower bands are only vectors or muggle 1 proteins.

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