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Hope
College Physics Department Research Experiences for Undergraduates Summer 2012 Project Summary |
| Project Title: | Stoichiometric Analysis of Metalloproteins in Thin Polyacrylamide Gels by Simultaneous Particle-Induced X-ray Emission Spectroscopy and Nuclear Reaction Analysis |
| Student Name: | Andrew McCubbin |
| Student's Home Institution: | Hope College |
| Research Advisor: | Dr. Paul DeYoung and Dr. Graham Peaslee |
| Source of Support: | This work is supported by the National Science Foundation under NSF-REU Grant No. PHY/DMR-1004811. |
Particle-Induced X-ray Emission Spectroscopy (PIXE) and Nuclear Reaction Analysis
(NRA) are two highly sensitive ion beam analytical techniques which probe the
elemental constitution and areal density, respectively, of a sample. Using
a 1.7 MV tandem electrostatic particle accelerator, these methods have been
utilized simultaneously to determine the identity and stoichiometry of metal
atoms within protein samples of known metal content; results indicate the technique
is a useful analytical tool which can be accurately extended to proteins of
hitherto unknown metal stoichiometries. The nondestructive technique, originally
developed for dried protein spots on a suitably thin support, has been extended
for metalloproteins suspended in polyacrylamide gel. This allows use of the
technique in biochemical applications in which proteins are separated by polyacrylamide
gel electrophoresis (PAGE), after which proteins are not easily removed from
the gel. Various methods of sample preparation such as sample mounting and
gel drying have been considered. Additionally, such factors as the thickness,
constitution, and possible contaminants of the gel have been investigated and
are reported.
Publications and Presentations:
“Ion Beam Analysis of Metalloprotein Stoichiometry”, Andrew McCubbin. 2011 Celebration of Undergraduate Research, Hope College, April 15, 2011.
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