Literature DB >> 23458356

Recorded scan times can limit the accuracy of sedimentation coefficients in analytical ultracentrifugation.

Huaying Zhao1, Rodolfo Ghirlando, Grzegorz Piszczek, Ute Curth, Chad A Brautigam, Peter Schuck.   

Abstract

We report systematic and large inaccuracies in the recorded elapsed time in data files from the analytical ultracentrifuge, leading to overestimates of the sedimentation coefficients of up to 10%. This far exceeds previously considered factors contributing to the uncertainty in this parameter and has significant ramifications for derived parameters such as hydrodynamic shape and molar mass estimates. The source of this error is currently unknown, but we found it to be quantitatively consistent across different instruments, increasing with rotor speed. Furthermore, its occurrence appears to correlate with the use of the latest data acquisition software from the manufacturer, in use in some of our laboratories for nearly 2 years. Many of the recently published sedimentation coefficients may need to be reexamined. The problem can be easily recognized by comparing the file timestamps provided by the operating system with the elapsed scan times recorded within the data files. Therefore, we implemented a routine in SEDFIT that can automatically examine the data files, alert the user to significant discrepancies, and correct the scan times accordingly. This eliminates errors in the recorded scan times. Published by Elsevier Inc.

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Year:  2013        PMID: 23458356      PMCID: PMC3676908          DOI: 10.1016/j.ab.2013.02.011

Source DB:  PubMed          Journal:  Anal Biochem        ISSN: 0003-2697            Impact factor:   3.365


  24 in total

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9.  The boundary structure in the analysis of reversibly interacting systems by sedimentation velocity.

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Review 10.  Evaluating the stoichiometry of macromolecular complexes using multisignal sedimentation velocity.

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  55 in total

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3.  Variable Field Analytical Ultracentrifugation: II. Gravitational Sweep Sedimentation Velocity.

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4.  Enhanced Sample Handling for Analytical Ultracentrifugation with 3D-Printed Centerpieces.

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7.  Solution properties of γ-crystallins: hydration of fish and mammal γ-crystallins.

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9.  Solution properties of γ-crystallins: compact structure and low frictional ratio are conserved properties of diverse γ-crystallins.

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10.  Improving the thermal, radial, and temporal accuracy of the analytical ultracentrifuge through external references.

Authors:  Rodolfo Ghirlando; Andrea Balbo; Grzegorz Piszczek; Patrick H Brown; Marc S Lewis; Chad A Brautigam; Peter Schuck; Huaying Zhao
Journal:  Anal Biochem       Date:  2013-05-24       Impact factor: 3.365

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