MAGP1

MAGP-1 and fibrillin-1, two protein components of extracellular microfibrils, were shown by immunoprecipitation studies to interact with the chondroitin sulfate proteoglycan decorin in the medium of cultured fetal bovine chondrocytes. Decorin interacted with each protein individually and with both proteins together to form a ternary complex. Expression of truncated fibrillin-1 proteins in Chinese hamster ovary cells localized proteoglycan binding to an amino-terminal region near the proline-rich domain. A spatially analogous fibrillin-2 truncated protein did not coprecipitate the same sulfated molecule, suggesting that chondroitin sulfate proteoglycan binding in this region is specific for fibrillin-1. An interaction between fibrillin and MAGP-1 was also observed under culture conditions that abrogated decorin secretion, suggesting that the two microfibrillar proteins can associate in the absence of the proteoglycan. Sulfation of matrix proteins is important for elastic fiber assembly because inhibition of sulfation was shown to prevent microfibrillar protein incorporation into the extracellular matrix of cultured cells.\n\nApplications:\nSuitable for Western Blot, ELISA, Immunoprecipitation, Immunohistochemistry and Immunocytochemistry. Other applications not tested.\n\nRecommended Dilution:\nELISA: 1:1000 -1:2000\nWestern Blot: 1:200 -1:500\nImmunoprecipitation: 1:100 -1:200\nImmunohistochemistry: 1:50 -1:100\nOptimal dilutions to be determined by the researcher.\n\nIn many cases, antigenicity of elastic fiber components can be enhanced by treating with 6M guanidine HCl, 50mM dithiothreitol, 20mM Tris pH 8.0 for 15 minutes. After washing with 20mM Tris pH 8.0, the slides are treated with 100mM iodoacetamide in the dark for 15 minutes, washed and stained using normal protocols.\n\nStorage and Stability:\nLyophilized powder may be stored at -20 degrees C. Stable for 12 months at -20 degrees C. Reconstitute by adding sterile 40-50% glycerol, ddH2O. Aliquot and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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