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Antibodies for denatured human H-ferritin stain only reticuloendothelial cells within the bone marrow
- Source :
- Scopus-Elsevier
-
Abstract
- Human H-ferritin homopolymer was denatured in sodium dodecyl sulphate and injected in mice to obtain antibodies for dissociated H-subunit. The antisera and Moabs obtained were specific for the denatured H-chain with no cross-reactivity with assembled ferritins in immunoblotting experiments. In contrast the Moabs for native recombinant H-ferritin are specific for the assembled ferritin molecules with weak cross-reactivity with the denatured H-subunits. The epitope recognized by one of the anti-denatured H-chain Moabs was mapped on the C-terminal helix of ferritin. The antibodies were used to study H-ferritin conformation in cells. In immunocytochemistry experiments the antibodies for denatured H-ferritin stained HeLa and K562 cells weakly, with a different intensity and pattern to those obtained with anti-native H-ferritin antibody. In human bone marrow smears the anti-denatured ferritin antibodies stained only reticuloendothelial cells, and did not recognize the H-ferritin rich immature erythroblasts. It is concluded that assembled and denatured H-ferritins are immunogenically distinct, and that erythroid and reticuloendothelial cells within the bone marrow contain H-ferritin in different conformations.
- Subjects :
- Protein Denaturation
Iron
Blotting, Western
Immunocytochemistry
Fluorescent Antibody Technique
Bone Marrow Cells
Biology
Epitope
law.invention
Bone Marrow
law
medicine
Humans
Denaturation (biochemistry)
Mononuclear Phagocyte System
Erythroid Precursor Cells
Antiserum
Staining and Labeling
Antibodies, Monoclonal
Hematology
Immunohistochemistry
Molecular biology
Ferritin
medicine.anatomical_structure
Ferritins
biology.protein
Recombinant DNA
Electrophoresis, Polyacrylamide Gel
Bone marrow
Antibody
HeLa Cells
Subjects
Details
- Database :
- OpenAIRE
- Journal :
- Scopus-Elsevier
- Accession number :
- edsair.doi.dedup.....1de0b4255ffcc64bceb69f9afcc50eed