Category: Plasma (Page 4)

Secreted Epididymal Glycoprotein: MATERIALS AND METHODS(2)

Immunocytochemical Analysis of Tissue Sections Small pieces of epididymal tissue were snap frozen in iso-pentane at — 80°C and embedded in Cryo-M-Bed (Bright) medium. Sections 8-12 l^m thick were cut on a cryostat, dried onto clean glass slides, and stored at —20°C until analysis. Dried sections were immersed in PBS containing 3% (w/v) BSA at room temperature for at least 1 h to block nonspecific…

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Secreted Epididymal Glycoprotein: MATERIALS AND METHODS(1)

Materials Unless otherwise stated, all routine chemicals were supplied by Sigma-Aldrich (Poole, UK) or Merck (Poole, UK). ‘‘Immobilon-P’’ PvDf membrane was supplied by Millipore (Watford, UK) and synthetic oligonucleotides by Sigma Genosys (Pampisford, UK). Culture supernatants containing McAbs to rat sperm membrane antigens 2D6 and 2B1, together with rabbit polyclonal antibodies (PAbs) to PBP and 2D6 antigens, were prepared and used as described previously. The…

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Secreted Epididymal Glycoprotein: INTRODUCTION(2)

Second, the specificity and tenacity with which epididymal glycoproteins bind to the sperm plasma membrane, and ultimately their structure within it, remain problematic. Some glycoproteins are readily dissociated from the sperm surface by high ionic strength solutions (e.g., ASF, HIS-50 ), others are only released after treatment with phospha-tidylinositol-specific phospholipase C indicating the presence of a GPI anchor (e.g., CD52 ), while most require detergents…

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Secreted Epididymal Glycoprotein: INTRODUCTION(1)

Mammalian spermatozoa acquire their motility and fertilizing capacity during passage through the epididymal duct. Concomitant with these changes in functionality, there is reorganization of the nucleoprotein; alterations in the structure and composition of the acrosome, mitochondria, and flagellum; and remodeling of the plasma membrane. The changes to the plasma membrane are of special significance, as it is primarily this organelle that regulates cross talk with…

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