Journal: Journal of Biological Chemistry

Article Title: The TspanC8 Subgroup of Tetraspanins Interacts with A Disintegrin and Metalloprotease 10 (ADAM10) and Regulates Its Maturation and Cell Surface Expression

doi: 10.1074/jbc.m112.416503

Figure Lengend Snippet: FIGURE 1. ADAM10 is a component of tetraspanin microdomains on platelets because ADAM10 immunoprecipitates from platelet lysates contain a characteristic pattern of tetraspanins and associated proteins. Human platelets were surface-biotinylated and lysed in 1% Brij97 lysis buffer (labeled B), which largely maintains tetraspanin-tetraspanin interactions and hence microdomain integrity, or in 1% Triton X-100 (labeled T), which dis- rupts tetraspanin microdomains (12). Lysates were immunoprecipitated (i.p) with negative control IgG or mAbs to ADAM10, tetraspanin CD151 (positive control), FcRIIa, or PECAM-1 (non-tetraspanin-associated negative control). Immunoprecipitates were analyzed by streptavidin Western blotting to detect biotinylated proteins. The separating line between the FcRIIa and PECAM-1 data indicates that the latter was from a different part of the same gel.

Article Snippet: The rat anti-mouse ADAM10 mAb was 139712 (R&D Systems), control mouse IgG1 was MOPC-21 (Sigma), and control rat IgG2A was 54447 (R&D Systems).

Techniques: Lysis, Labeling, Immunoprecipitation, Negative Control, Positive Control, Western Blot