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Purification of three Morgue-associated proteins.

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posted on 30.09.2013, 01:56 by Ying Zhou, Yiqin Wang, Barbara A. Schreader, John R. Nambu

A. Silver staining of an analytical SDS polyacrylamide gel separating proteins from whole fly extracts that associate with Morgue. Protein bands corresponding to Morgue-3xFLAG (∼60 kD) (black arrowhead) as well as the anti-FLAG Ig heavy chain (∼50 kD) are indicated (gray arrowhead). Lanes 1–4 correspond to material purified via the anti-FLAG resin from the following flies: Lane 1: P[da-Gal4]; Lane 2: P[UAS-Morgue3xFlag]; Lane 3: P[da-Gal4], P[UAS-3xFlag:Morgue]; Lane 4: P[da-Gal4], P[UAS-Morgue3xFlag]; Lane 5: Protein molecular weight markers. The three major bands corresponding to Morgue-associated proteins purified are indicated (arrows) and correspond to polypeptides migrating between 28 kD and 20 kD. B. Coomassie Brilliant Blue staining of a preparatory SDS polyacrylamide gel separating Morgue-associated proteins from extracts of P[da-Gal4], P[UAS-Morgue:3xFlag] adult flies purified via anti-FLAG resin. Three major bands (arrows) were excised from the gel and the corresponding polypeptides analyzed via mass spectrometry. Lane on right corresponds to protein molecular weight markers.


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