Optimisation of a laser ablation cell for detection of hetero-elements in proteins blotted onto membranes by use of inductively coupled plasma mass spectrometry

Feldmann, Ingo; Koehler, Christina U.; Roos, Peter H.; Jakubowski, Norbert

doi:10.1039/b606773e

Cited by 63 publications

(60 citation statements)

References 31 publications

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“…Therefore some form of post-separation treatment of the gel is required. In most published studies it is mentioned that the gel was dried before ablation, although no details are given [6,24,25,34].…”

Section: Preparation Of Gel For Lamentioning

confidence: 99%

Evaluation of gel electrophoresis conditions for the separation of metal‐tagged proteins with subsequent laser ablation ICP‐MS detection

et al. 2009

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Although laser ablation (LA)-ICP-MS has been reported for the determination of metalloproteins separated by gel electrophoretic techniques (GE), systematic studies that define the conditions essential for successful measurements are still scarce. In this paper we present the results of our studies of basic conditions for the effective application of GE-LA-ICP-MS for the separation of metal-binding proteins, focusing on their stability during GE and post-separation gel treatment. The stability of metal-protein complexes (haemoglobin, myoglobin, superoxide dismutase, carbonic anhydrase, transferrin, albumin, cytochrome c) during GE is dependent on the nature of the metal-protein interaction and the principle of separation. We have observed that non-denaturing GE is a suitable separation technique for most metal-protein complexes (e.g. Zn in carbonic anhydrase and Fe in Tf and myoglobin were quantitatively recovered in a spiked liver cytosol), whereas separation by denaturing GE strongly impaired the stability of the complexes. Equally important is the post-separation treatment of the gel to enable successful detection of the metal. LA-ICP-MS requires drying of the gel without loss of protein-bound metal or cracking of the gel. This was successfully achieved using glycerol followed by heating. We demonstrate that staining of the gel prior to LA-ICP-MS using silver or Coomassie blue is not recommended, since most protein-bound metal is lost during the staining procedure. Furthermore it has been shown that only line scanning with a speed of less than 30 microm/s can reliably distinguish between lines 1 mm apart, while raster spot analysis carries the risk of misinterpretation due to contamination in/on inhomogeneous gels.

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Section: Preparation Of Gel For Lamentioning

confidence: 99%

Evaluation of gel electrophoresis conditions for the separation of metal‐tagged proteins with subsequent laser ablation ICP‐MS detection

et al. 2009

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“…The cell geometry is critical to performance, particularly the shape and volume, and the relative position of the inlet and outlet ports in relation to the sampling region. [30][31][32] Horstwood et al described a tear-drop shaped single volume cell (inner volume ca 3 cm 3 ) 33 , based on theoretical and experimental observations of particle transport by Bleiner and Günther 22 , which was capable of resolving single shots in 0.8 s. A limitation of the single volume design is that the response is not uniform across the cell surface. Furthermore, the very low inner volumes required for fast washout place significant restrictions on sample size.…”

Section: Introductionmentioning

confidence: 99%

High-Speed, Integrated Ablation Cell and Dual Concentric Injector Plasma Torch for Laser Ablation-Inductively Coupled Plasma Mass Spectrometry

et al. 2015

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In recent years, laser ablation-inductively coupled plasma mass spectrometry (LA-ICPMS) has gained increasing importance for biological analysis, where ultratrace imaging at micrometer resolution is required. However, while undoubtedly a valuable research tool, the washout times and sensitivity of current technology have restricted its routine and clinical application. Long periods between sampling points are required to maintain adequate spatial resolution. Additionally, temporal signal dispersion reduces the signal-to-noise ratio, which is a particular concern when analyzing discrete samples, such as individual particles or cells. This paper describes a novel, two-volume laser ablation cell and integrated ICP torch designed to minimize aerosol dispersion for fast, efficient sample transport. The holistic design utilizes a short, continuous diameter fused silica conduit, which extends from the point of ablation, through the ICP torch, and into the base of the plasma. This arrangement removes the requirement for a dispersive component for argon addition, and helps to keep the sample on axis with the ICP cone orifice. Hence, deposition of sample on the cones is theoretically reduced with a resulting improvement in the absolute sensitivity (counts per unit mole). The system described here achieved washouts of 1.5, 3.2, and 4.9 ms for NIST 612 glass, at full width half, 10%, and 1% maximum, respectively, with an 8–14-fold improvement in absolute sensitivity, compared to a single volume ablation cell. To illustrate the benefits of this performance, the system was applied to a contemporary bioanalytical challenge, specifically the analysis of individual biological cells, demonstrating similar improvements in performance.

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“…364 For this purpose a particular laser ablation cell was designed. 365 Antibodies were labelled specifically at lysine residues by means of lanthanidecontaining isothiocyanato benzyl-(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) (DOTA) 366 or by iodination 367 which preferentially occurs at the two ortho-positions of the hydroxyl group of tyrosine.…”

Section: Speciation Analysismentioning

confidence: 99%

Inductively coupled plasma- and glow discharge plasma-sector field mass spectrometry : Part II. Applications

Jakubowski

Prohaska

Vanhaecke

et al. 2011

J. Anal. At. Spectrom.

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Optimisation of a laser ablation cell for detection of hetero-elements in proteins blotted onto membranes by use of inductively coupled plasma mass spectrometry

Cited by 63 publications

References 31 publications

Evaluation of gel electrophoresis conditions for the separation of metal‐tagged proteins with subsequent laser ablation ICP‐MS detection

Evaluation of gel electrophoresis conditions for the separation of metal‐tagged proteins with subsequent laser ablation ICP‐MS detection

High-Speed, Integrated Ablation Cell and Dual Concentric Injector Plasma Torch for Laser Ablation-Inductively Coupled Plasma Mass Spectrometry

Inductively coupled plasma- and glow discharge plasma-sector field mass spectrometry : Part II. Applications

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