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. 2005 May 10;388(Pt 1):195–204. doi: 10.1042/BJ20041797

Figure 6. Co-sedimentation of and co-immunoprecipitation of STNB and DAP-160.

Figure 6

(A) A 5–25% glycerol gradient on which was loaded S1 (1000 g supernatant) from homogenized wild-type fly heads. Fifteen fractions were collected, and each fraction run on SDS/PAGE (7% gel), Western blotted and probed separately with antibodies against STNB, DAP-160, dynamin (DYN) and SYT-I. Fraction 1 represents the top and fraction 15 the bottom of the gradient. The peak of SYT-I in fractions 5 through 8 represents free synaptic vesicles [15]. STNB co-sediments with this fraction, whereas DAP-160, and to a lesser extent dynamin, is enriched in these fractions. (B) A Triton X-100-solubilized P3 fraction (see the Experimental section) contains both STNB and DAP-160. Addition of anti-STNB antibodies, followed by Protein A–Sepharose to this fraction, precipitates both STNB and DAP-160. The amount of the immunoprecipitated fraction loaded on the gel is equivalent in volume to that shown in the P3 fraction. Whereas most of the STNB protein is immunopreciptated from the P3 fraction, only a small amount of the total DAP-160 is co-precipitated.