Isolation of laminin from human placental basement membranes: amnion, chorion and chorionic microvessels.
Laminin components were solubilized from basement membranes of amnion, chorion and chorionic microvessels of human placenta without prior protease digestion. These structures, after isolation, were initially processed in a sonicator bath containing a solution of Triton X-100, EDTA and 2M NaCl and the laminins extracted sequentially with 0.5M NaCl, 8 M urea, and 8 M urea + 2% 2-mercaptoethanol + 2% SDS. A high molecular weight (appr. 1 x 10(6)) complex containing laminins was purified by gel filtration on a Sepharose CL-2B column. This complex migrated as a single band on gel electrophoresis before reduction but resolved, after reduction, into four major laminin components, laminin A (350,000 M.W.), laminin M (240,000 M.W.) and laminins B1 and B2 (195,000 and 185,000 M.W.). Laminin M is a new molecular species of this protein.
D000596:Amino Acids / Q000302:isolation & purification; D000650:Amnion / Q000032:analysis*; D000818:Animals; D001485:Basement Membrane / Q000032:analysis; D055598:Chemical Phenomena; D002621:Chemistry; D002823:Chorion / Q000032:analysis* / Q000098:blood supply; D005260:Female; D006023:Glycoproteins / Q000302:isolation & purification* / Q000502:physiology; D006801:Humans; D007120:Immunochemistry; D007797:Laminin; D008565:Membrane Proteins / Q000302:isolation & purification*; D008833:Microcirculation / Q000032:analysis; D010920:Placenta / Q000032:analysis*; D011247:Pregnancy; D051381:Rats; D012995:Solubility
D000596:Amino Acids; D006023:Glycoproteins; D007797:Laminin; D008565:Membrane Proteins