Issue 7, 2014

Development of a calibration and standardization procedure for LA-ICP-MS using a conventional ink-jet printer for quantification of proteins in electro- and Western-blot assays

Abstract

We developed new procedures for internal standardization and calibration to be used for laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) for elemental micro mapping imaging of biological samples like Western blot membranes and tissue sections. These procedures are based on printing of metal spiked inks onto the top of thin layer samples for simultaneous internal standardization and calibration of LA-ICP-MS. In the case of internal standardization the ink is spiked with indium as an internal standard and homogenously printed over the entire membrane (size 56 cm2) prior to LA-ICP-MS detection, a standard deviation (RSD) value of 2% was achieved. In the second approach the metal content of lanthanide tagged proteins and antibodies after biological work flows was quantified by LA-ICP-MS on nitro-cellulose membranes. In this case the inks spiked with varying metals were printed with different densities on the same nitrocellulose membranes in well-defined squares to produce matrix-matched calibration standards. For validation and calibration the ink squares were excised and the specific metal content was measured by liquid ICP-MS after solubilization of the membrane slice. For the printed calibration standard limits of detection (LOD) of <4 fmol for different metals and relative process standard deviations of 1–2% only were determined via LA-ICP-MS.

Graphical abstract: Development of a calibration and standardization procedure for LA-ICP-MS using a conventional ink-jet printer for quantification of proteins in electro- and Western-blot assays

Supplementary files

Article information

Article type
Paper
Submitted
13 二月 2014
Accepted
02 四月 2014
First published
02 四月 2014

J. Anal. At. Spectrom., 2014,29, 1282-1291

Development of a calibration and standardization procedure for LA-ICP-MS using a conventional ink-jet printer for quantification of proteins in electro- and Western-blot assays

S. Hoesl, B. Neumann, S. Techritz, M. Linscheid, F. Theuring, C. Scheler, N. Jakubowski and L. Mueller, J. Anal. At. Spectrom., 2014, 29, 1282 DOI: 10.1039/C4JA00060A

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements