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  • Antigen  (1)
  • basic fibroblast growth factor  (1)
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Methods in cell science 10 (1986), S. 125-132 
    ISSN: 1573-0603
    Keywords: basic fibroblast growth factor ; heparin affinity ; hepatoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Intact human hepatoma derived basic fibroblast growth factor (bFGF) is a cationic 18 400-molecular-weight polypeptide, which stimulates the proliferation of 3T3 and endothelial cells at 1 ng/ml. bFGF has a strong affinity for heparin, and can be purified to homogeneity from human hepatoma cells using heparin-Sepharose chromatography. A three-step procedure is used: (a) extraction of the cells with 1M NaCl at pH 7.5; (b) Bio-Rex 70 cation exchange chromatography; and (c) heparin-Sepharose chromatography. The molecular weight of bFGF depends on the pH of the cell extraction step. When extracted at neutral pH, intact bFGF is obtained with a molecular weight of 18 400, however when extracted at pH 3.5 to 4.5 the molecular weight of bFGF is about 16 500. Lowering the molecular weight by acid pH extraction can be inhibited with a mixture of the proteinase inhibitors, PMSF, leupeptin, and pepstatin. These results suggest that degradation of bFGF is the result of cleavages by acid proteinases. It has been determined by amino acid sequence data and western blot analysis that the cleavages occur at the amino terminus of bFGF. The lower molecular weight form of bFGF has the same biological activity and exhibits the same chromatographic behavior on heparin-Sepharose as does intact bFGF.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 41 (1995), S. 249-256 
    ISSN: 1040-452X
    Keywords: Spermatozoa ; Protein ; Antigen ; Fertilization ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The binding of the spermatozoon to the zona pellucida is a species-specific phenomenon. We have previously shown that the binding of hamster sperm to the homologous zona pellucida involves a sperm 26-kDa glycoprotein, the P26h, originating in the epididymis. In order to establish to what extent this sperm protein is involved in the species-specific recognition of the egg's extracellular coat, we have compared the inhibitory properties of anti-P26h antibodies in a sperm-zona pellucida assay using hamster and mouse gametes. Anti-P26h IgGs inhibit, in a dose-dependent manner, gamete interactions in both species, although in a less efficient manner in the mouse than in the hamster. While anti-26kDa Fab fragments are as efficient as the intact IgG to inhibit hamster sperm-zona pellucida binding, they have no effect on mouse gamete interaction. ELISA, Western blot, and immunohistochemical experiments have been performed in order to characterize the mouse antigen(s) recognized by the anti-P26h antiserum. ELISA and Western blots showed that this antiserum recognized two proteins on mouse spermatozoa that are less reactive than the hamster P26h. These antigens are localized in the acrosomal region of epididymal spermatozoa of both species. These results indicate that the hamster P26H involved in zona pellucida interaction has certain unique epitopes, while others are common to the sperm of both species. © 1995 Wiley-Liss, Inc.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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