CIL:12860, Rattus rattus, glandular epithelial cell, milk secreting cell, mammary alveolar cell. In Cell Image Library ...

• Main Authors: Ladinsky, Mark, Howell, Kathryn E.
• Format: Dataset
• Language: unknown
• Published: UC San Diego Library Digital Collections 2021
• Subjects: secretion; milk secreting cell; glandular epithelial cell; smooth endoplasmic reticulum; lipid storage; lipid particle; trans-Golgi network; mammary alveolar cell; protein binding; Mitochondrion; lactation; Rattus rattus
• Online Access: https://dx.doi.org/10.6075/j0hq3z2z; https://library.ucsd.edu/dc/object/bb6457743f

Immunoelectron microscopy of mammary gland tissue from lactating rats with the trans-Golgi network identified by immunogold labeling of TGN38, a transmembrane trans-golgi protein. Note that trans‐Golgi cisternae in lactating mammary cells contain large amounts of casein and other milk proteins and are grossly expanded due to the osmolarity of the secreted fluid. ... : Tissue was processed for immunoelectron microscopy using a modified Tokuyasu method. Briefly, minced tissue was fixed in 4% paraformaldehyde containing 5% sucrose and 100 mM HEPES and infiltrated with PBS containing 2.1 M sucrose over 10 h, with repeated solution changes. Fixed tissue was transferred to an aluminum cryosectioning stub (Ted Pella, Inc., Redding, CA) and immediately frozen in liquid nitrogen. Semithin (90 nm) cryosections were cut at -110C with an UltraCut UCT/FCS cryomicrotome (Leica), using a diamond knife (Diatome) and transferred to a Formvar-coated, carbon-coated, glow-discharged 100-mesh copper-rhodium electron microscopy grid. Following blocking of nonspecific antibody binding sites with 10% calf serum in PBS, the sections were labeled by incubation with an antibody to TGN38 (2F7, mouse monoclonal) followed by colloidal gold-conjugated secondary antibody (10 nm anti-mouse; Ted Pella Inc., Redding, CA) and then negatively stained and embedded with 1% uranyl acetate, 1% methylcellulose in ...