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. 1999 May 15;13(10):1289–1296. doi: 10.1101/gad.13.10.1289

Figure 2.

Figure 2

(A) Immunoblot analysis of six-subunit Drosophila ORC using antibodies raised against individual ORC subunits. Protein samples (∼20 ng of total ORC estimated by silver-stained material in the samples) corresponding to purified Drosophila embryonic ORC and immunoprecipitated by α-ORC2 or α-ORC6 antibodies, material was separated using SDS-PAGE and transferred to a PVDF membrane. Membrane strips were incubated with antibodies raised against individual Drosophila ORC subunits as indicated by the side labels. Signals with anti-ORC3 and anti-ORC6 sera were obtained first and the same filter was subsequently probed with the ORC4 reagent. Low levels of specific ORC1 and ORC2 proteolytic fragments are also visible. Protein bands were visualized by subsequent ECL assay. (B) Silver-stained gel of Drosophila ORC immunoprecipitated by α-ORC2 (lane 1) and recombinant baculovirus-expressed Drosophila ORC as eluted from a Ni column (lane 2) and subsequently purified on a glycerol gradient (lane 3). Lanes 1 and 2 are taken from the same gel. DmORC1 has an altered electophoretic mobility due to addition of a His tag.