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Antigenic Protein In Microgravity-Grown Human Mixed Mullerian Tumor (LN1) Cells Preserved In RNA Stabilizing AgentCells treated with RNAlater(TradeMark) have previously been shown to contain antigenic proteins that can be visualized using Western blot analysis. These proteins seem to be stable for several months when stored in RNA stabilizer at 4 C. Antigenic protein can be recovered from cells that have been processed using an Ambion RNAqueous(Registered TradeMark) kit to remove RNA. In this set of experiments, human mixed Mullerian tumor (LNI) cells grown on the International Space Station during Expedition 3 were examined for antigenic stability after removal of RNA. The cells were stored for three months in RNAlater(TradeMark) and RNA was extracted. The RNA filtrate containing the protein was precipitated, washed, and suspended in buffer containing sodium dodecyl sulfate (SDS). Samples containing equal concentrations of protein were loaded onto SDS-polyacrylamide gels. Proteins were separated by electrophoresis and transferred by Western blot to polyvinylidene fluoride (PVDF) membrane. The Western blots were stained with an enhanced chemiluminescent ECL(Registered Trademark) Plus detection kit (Amersham) and scanned using a Storm 840 gel image analyzer (Amersham, Molecular Dynamics). ImageQuant(Registered TradeMark) software was used to quantify the densities of the protein bands. The ground control and flight LN1 cell samples showed a similar staining pattern over time with antibodies to vimentin, glyceraldehyde-3-phosphate dehydrogenase, and epithelial membrane antigens.
Document ID
20060019175
Acquisition Source
Johnson Space Center
Document Type
Conference Paper
Authors
Hammond, Dianne K.
(Enterprise Advisory Services, Inc. Houston, TX, United States)
Becker, Jeanne
(National Space Biomedical Research Inst. Houston, TX, United States)
Elliott, T. F.
(Wyle Life Sciences, Inc. Houston, TX, United States)
Holubec, K.
(Wyle Life Sciences, Inc. Houston, TX, United States)
Baker, T. L.
(Wyle Life Sciences, Inc. Houston, TX, United States)
Love, J. E.
(NASA Johnson Space Center Houston, TX, United States)
Date Acquired
August 23, 2013
Publication Date
January 1, 2004
Subject Category
Life Sciences (General)
Meeting Information
Meeting: 2004 ASGSB Meeting
Location: Brooklyn, NY
Country: United States
Start Date: November 9, 2004
End Date: November 12, 2004
Sponsors: American Society for Gravitational and Space Biology
Funding Number(s)
CONTRACT_GRANT: NAS9-1341
CONTRACT_GRANT: NAG9-1341
Distribution Limits
Public
Copyright
Work of the US Gov. Public Use Permitted.

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