Isolation and characterization of endosomes from rat liver
Thesis/Dissertation
·
OSTI ID:6905389
Three fractions of rat liver endosomes, called 50 Kg Light, 50 Kg Heavy, and 150 Kg have been isolated on 16% Percoll gradients. The 50 Kg Heavy fraction accumulates ligand as a function of time after injection, using either /sup 125/I-asialoorosomucoid (/sup 125/I-ASOR) or /sup 125/I-immunoglobulin A (/sup 125/I-IgA) as ligands. A pulse-chase protocol was also used to study the kinetics of ligand entry into the endosomal compartments. A double-label, 3,3'-diaminobenzidine (DAB)-induced density shift protocol was used to study the internalization of two ligands with different destinations in the hepatocyte. Rats were injected intraportally with /sup 125/I-ASOR-HRP and /sup 131/I-IgA and the liver was fractionated at various times post-injection. The three ligand-containing endosomal fractions were isolated and each subjected to the DAB shift procedure. This treatment causes organelles containing /sup 125/I-ASOR-HRP and another ligand occupying the same compartment to shift to a higher density. Thus, information on whether the /sup 131/I-IgA is colocalized or segregated from the /sup 125/I-ASOR-HRP can be obtained. The authors have used an instantaneous pulse, temperature shift protocol to study the heterogeneity of these three endosomal fractions isolated from rat liver.
- Research Organization:
- Case Western Reserve Univ., Cleveland, OH (USA)
- OSTI ID:
- 6905389
- Country of Publication:
- United States
- Language:
- English
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OSTI ID:6024530
Related Subjects
550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BODY
CARBOHYDRATES
COMPARTMENTS
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
ELECTRON CAPTURE RADIOISOTOPES
GLANDS
GLOBULINS
IMMUNOGLOBULINS
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE 131
IODINE ISOTOPES
ISOTOPES
KINETICS
LABELLED COMPOUNDS
LIGANDS
LIVER
LIVER CELLS
MAMMALS
MUCOPROTEINS
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
POLYSACCHARIDES
PROTEINS
RADIOISOTOPES
RATS
REACTION KINETICS
RODENTS
SACCHARIDES
SOMATIC CELLS
UPTAKE
VERTEBRATES
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BODY
CARBOHYDRATES
COMPARTMENTS
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
ELECTRON CAPTURE RADIOISOTOPES
GLANDS
GLOBULINS
IMMUNOGLOBULINS
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE 131
IODINE ISOTOPES
ISOTOPES
KINETICS
LABELLED COMPOUNDS
LIGANDS
LIVER
LIVER CELLS
MAMMALS
MUCOPROTEINS
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
POLYSACCHARIDES
PROTEINS
RADIOISOTOPES
RATS
REACTION KINETICS
RODENTS
SACCHARIDES
SOMATIC CELLS
UPTAKE
VERTEBRATES