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. 1997 Aug 11;138(3):719–728. doi: 10.1083/jcb.138.3.719

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Western blot analysis of the laminin 5–type VII collagen–NC-1 complex immunoaffinity purified from collagenase extracts of amniotic membranes. Type VII collagen–NC-1 was first affinity purified on mAb NP-32 from collagenase extracts of amniotic membranes. Eluted fractions of NC-1 were pooled, dialyzed against PBS, and passed over a mAb 6F12 affinity chromatography column. Bound materials were immunoblotted with polyclonal anti-laminin 5 (pLm5; lane 3), and anti-NC-1 (mAb NP-32, lane 4). The bound material shows the pattern obtained for purified laminin 5 (lane 1), which does not contain NC-1 (lane 2) and for purified NC-1 (not shown, but identical to lane 6). The nonbound fraction contains NC-1 only (lane 6) but no laminin 5 (lane 2).