Cloning and functional verification of a full-length ASH1 cDNA. (A) Map of Drosophila ASH1 protein indicating known domains [SwissProt:Q9VW15]. Subcloned RT-PCR amplicons used for cDNA assembly are shown below. (B) ASH1-specific antibodies, raised against epitopes residing in the N- and C-terminal portion of the protein, detect ASH1-GFP fusion products obtained by transfecting Drosophila cells with expression constructs stated on top. (C) Sanger sequencing trace indicating missing amino acids T1716-L1717 in cloned ASH1 cDNA. (D) Comparision of target and cross reactivity between the ASH1-specific antibodies ASH1-C and Q4177. Immunoblots of S2-DRSC derived whole cell (Cell) and nuclear lysate (Nuc.) were prepared using the antibody dilutions given on top.
Kockmann et al. Genome Biology 2013 14:R18 doi:10.1186/gb-2013-14-2-r18