Abstract
Iron in blood plasma is bound to its transport protein transferrin, which delivers iron to most tissues. In iron overload and certain pathological conditions, the carrying capacity of transferrin can become exceeded, giving rise to non-transferrin-bound iron, which is taken up preferentially by the liver, kidney, pancreas, and heart. The measurement of tissue transferrin- and non-transferrin-bound iron (TBI and NTBI, respectively) uptake in vivo can be achieved via intravenous administration of 59Fe-labeled TBI or NTBI followed by gamma counting of various organs. Here we describe a detailed protocol for the measurement of TBI and NTBI uptake by mouse tissues.
Keywords: NTBI, TBI, 59Fe, Tissue iron uptake, Iron overload
Materials and Reagents
Equipment
Procedure
Notes
After each experiment, use radioactive cleaning reagent to clean the surgical tools, mouse cages, and related equipment to decontaminate 59Fe. Perform a radioactive swipe test on the related equipment and area where the experiment is performed after each experiment to ensure no contamination is observed. Radioactive waste products should be disposed of by following your local Environmental Health and Safety (EH&S) regulations.
Recipes
Acknowledgments
This work was supported by the National Institutes of Health grant DK080706 (to M.D.K.). The protocol was adapted from Craven et al. (1987), and a short version of the adapted protocol was published in Jenkitkasemwong et al. (2015).
References
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