Abstract
Fibronectin, a high-molecular glycoprotein of the extracellular matrix and plasma, has provoked increasing interest in ophthalmology because of its possibly important role in the pathogenesis of proliferative vitreoretinopathy (PVR). A quantitative determination of fibronectin in vitreous aspirates is necessary for the evaluation of this glycoprotein in PVR. A noncompetitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantification of immunoreactive fibronectin in human plasma samples and vitreous as pirates. Here, procedures to establish an ELISA for the clinical laboratory routine are discussed and possible pitfalls during the assay standardization are described.
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Abbreviations
- ELISA:
-
enzyme-linked immunosorbent assay
- PBS:
-
phosphate buffered saline
- BSA:
-
bovine serum albumin
- AP:
-
alkaline phosphatase
- OD:
-
optical density
- SDS-PAGE:
-
sodium dodecylsulfate polyacrylamide gel electrophoresis
- PMSF:
-
phenylmethylsulfonyl fluoride
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Weller, M., Wiedemann, P., Heimann, K. et al. Fibronectin quantification in plasma and vitreous by a noncompetitive ELISA technique. Doc Ophthalmol 69, 341–351 (1988). https://doi.org/10.1007/BF00162748
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DOI: https://doi.org/10.1007/BF00162748