Figure 2 : In vitro binding assays using recombinant Drosophila Sarcoglycans.

(A) Diagrams of the obtained constructs for the production of recombinant proteins. The numbers written below the rectangular bars represent the amino acid numbers counted from N-terminus of respective proteins. (B) Western blots of both Input and Elution samples with anti-His or anti-Flag antibodies are shown. Binding between dScgα and dScgβ (I), dScgα and dScgδ (II), dScgβ and dScgδ (III) was tested. Binding between dScgα and lamin was also examined as a negative control to exclude the possibility of non-specific binding (IV). 5 mg of total protein in the E. coli extracts was applied for a 100 μl volume of the Ni-NTA agarose gel and eluted with 100 μl buffer. 20 μl of each sample was loaded for SDS-PAGE.

Yamaguchi et al.HOAJ BIOLOGY  2012 1:7DOI : 10.7243/2050-0874-1-7