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Figure 7


Fig. 7. Co-immunoprecipitation of ß5 integrin with Sdc1 requires the Sdc1 ectodomain. Western blots probed with rabbit polyclonal ß5 integrin (A,B), pan-syndecan or S1ED (C) antibody for detection of ß5 integrin and Sdc1, respectively, in immune complexes isolated after immunoprecipitation of full-length mouse Sdc1 (mAb 281.2), human Sdc1 constructs (mAb B-B4) and FcRecto-hS1 chimera (mAb 10.1) from pre-cleared B82L whole-cell lysates. In S1ED-competition experiments (A), 30 µM soluble GST, GST-mS1ED (with mAb B-B4) or GST-hS1ED (with mAb 281.2) was added to the reaction mixture. Provided as a reference is a methanol precipitation (MeOH) of 300 µg of whole-cell lysate. ß5 integrin immunoblotting reveals a 110 kDa band, under reduced conditions, detectable in the mouse Sdc1 and select human Sdc1 (hS1, pLTM, {Delta}cyto), but not FcRecto-hS1 chimera immunoprecipitates or immunoprecipitates isolated with antibody isotype control IgG: mouse IgG1 (mIgG) for mAbs B-B4 and 10.1 and rat IgG2A (rIgG) for mAb 281.2.





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