Preparation of Placental (Fetal Tissue) Fibronectin and its Carbohydrates
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.
Publication Source (Journal or Book title)
Methods in Enzymology
Laine, R., Fisher, S., & Zhu, B. (1987). Preparation of Placental (Fetal Tissue) Fibronectin and its Carbohydrates. Methods in Enzymology, 144 (C), 420-429. https://doi.org/10.1016/0076-6879(87)44191-8