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. 2012 Dec 26;7(12):e52793. doi: 10.1371/journal.pone.0052793

Figure 7. WARP co-precipitates with collagen VI.

Figure 7

Collagen VI was isolated from cartilage extracts using magnetic beads. Following magnetic bead separation, total lysate, supernatant and pellet fractions were immunoblotted using antibodies against WARP and collagen VI. Fractions containing crude isolate (lanes 3 and 6), magnetic bead-isolated collagen VI (lanes 1 and 4), and supernatant remaining following collagen VI precipitation (lanes 2 and 5) were subjected to SDS-PAGE and immunoblotted using sheep anti-WARP antiserum (lanes 1 to 3) or anti-collagen VI antibodies (lane 4 to 6). WARP was present in collagen VI-precipitated material but not the supernatant indicating that both WARP and collagen VI are present in suprastructures. A control demonstrating that virtually no material is isolated by normal mouse serum coupled to magnetic beads is shown in lane 7 (total protein stain of blot membrane). Under reducing conditions WARP migrates as a 50 kDa monomer and the α1 and α2 chains of collagen VI co-migrate at approximately 200 kDa. Molecular weight marker (kDa) is shown on left. bp, bead pellet; sub, supernatants; crude, crude extract.