Separation of cellular iron containing compounds by electrophoresis
Language English Country United States Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
9533565
DOI
10.1007/bf02789087
Knihovny.cz E-resources
- MeSH
- Autoradiography MeSH
- Iron Chelating Agents chemistry MeSH
- Deferoxamine chemistry MeSH
- Electrophoresis, Polyacrylamide Gel MeSH
- Mice, Inbred BALB C MeSH
- Mice MeSH
- Sensitivity and Specificity MeSH
- Ultrafiltration MeSH
- Iron chemistry isolation & purification MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Iron Chelating Agents MeSH
- Deferoxamine MeSH
- Iron MeSH
High resolution separation of metalloproteins and other iron compounds based on native gel electrophoresis followed by 59Fe autoradiography is described. Lysates of mouse spleen erythroid cells metabolically labeled with 59Fe-transferrin were separated on 3-20% polyacrylamide gradient gels in the presence of Triton X100 and detected by autoradiography. In addition to ferritin and hemoglobin, several compounds characterized by their binding of iron under different conditions were described. Iron chelatable by desferrioxamine migrated in the region where several high-molecular weight compounds were detected by silver staining. The technique is nondissociative, allowing identification of iron compounds with the use of specific antibodies. Cellular iron transport and the action of iron chelators on specific cellular targets can be investigated in many small biological samples in parallel.
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