Secreted Epididymal Glycoprotein: MATERIALS AND METHODS(3)

METHODS(3)

Frozen sections were cut at —20°C using a Reichert FC4 cryo-ultramicrotome and incubated in undiluted 2D6 McAb supernatant for 3-5 h at 4°C. Control sections were incubated in culture medium only. After thorough washing in PBS/0.1% (w/v) BSA, sections were incubated in rabbit anti-mouse IgG diluted 1:1000 for 30 min followed by colloidal gold (5 or 15 nm)-conjugated goat anti-rabbit IgG for 1 h. After further washing, sections were counterstained with 2% (w/v) uranyl acetate, embedded in 0.3% (w/v) methyl cellulose, and air dried. Sections were viewed using a JOEL 100C transmission electron microscope as described. Flovent oral aerosol

Collection of Luminal Contents from the Caput and Cauda Epididymidis

Luminal contents were collected from the distal caput and cauda epi-didymidis either by micropuncture or by slicing the whole tissue. In the latter procedure, the epididymis was cleaned of excess blood and superficial fluid by gently blotting onto Kleenex paper and immersed in 2 ml PBS containing 1 mM Pefabloc. Three to four incisions were made with fine scissors, and luminal fluid was allowed to exude from the cut ends of the duct.

Category: Plasma

Tags: epididymis, gamete biology, male reproductive tract, sperm