Jun 30, 2025
Intracellular iron measurements by ICP-MS
- 1The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK.;
- 2Department of Clinical and Movement Neurosciences, UCL Queen Square Institute of Neurology, Queen Square, London, WC1N 3BG, UK.
Protocol Citation: Laura Mahoney-Sanchez 2025. Intracellular iron measurements by ICP-MS. protocols.io https://dx.doi.org/10.17504/protocols.io.j8nlkyb41g5r/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 30, 2025
Last Modified: June 30, 2025
Protocol Integer ID: 221273
Keywords: intracellular iron measurements by icp, intracellular iron measurement, mass spectrometry, icp
Abstract
This protocol describes intracellular iron measurements performed by Inductively coupled Plasma-Mass spectrometry.
Cell lysis
Cells are pelleted by centrifugation 800 x g, 20°C for 00:05:00
Supernatant is removed and cell pellet lysed by adding 250 µL of RIPA buffer + 1% Protease and phosphatase inhibitors.
Protein concentration was measured by Pierce BCA assay
Inductively Coupled Plasma-Mass Spectrometer
Cell lysis is combined with 0.5 mL of trace metal-grade concentrated HNO3 (67–69% w/w)
Samples are heated at 50 °C for 01:00:00
Sample digests are transferred into trace metal-grade 15ml centrifuge tubes and diluted with 4.45 mL of purified water from a Milli-Q system
Samples are spiked with50 µL of 5µg L-1 gallium
Iron concentrations are determined using a Perkin Elmer NexION 5000 Inductively Coupled Plasma-Mass Spectrometer operating in dynamic reaction cell mode with ammonia gas.
Normalise iron concentrations to total protein.