Preparation of Placental (Fetal Tissue) Fibronectin and its Carbohydrates

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This chapter describes the method used in laboratory for the preparation of human placental (fetal) tissue fibronectin. A major advantage of this technique is the relatively large amount of cell- and basement membrane-associated fibronectin which can be rapidly isolated: 50 to 120 mg of purified glycoprotein is purified from a single term placenta. Methods for isolating fibronectins are based on the affinity of all the glycoproteins for denatured collagen. Thus, the first step toward purification is usually chromatography on gelatin-Sepharose. Gel permeation chromatography of the bound fraction results in a relatively homogeneous protein preparation as demonstrated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The cost of purchasing sufficient heparin and DTT to extract additional fibronectin from the urea-insoluble residue from one placenta is about $200. If fibronectin is isolated on a routine basis, a simple, economic alternative is to process additional placentas. Because 50 mg of fibronectin can be extracted from only one placenta using urea alone the procedure can easily be “scaled-up” by the processing of several placentas simultaneously. © 1987, Elsevier Inc. All rights reserved.

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Methods in Enzymology

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