1997 Fiscal Year Final Research Report Summary
ESTABLISHMENT OF A NEW METHOD FOR PROTEIN ANALYSIS AFTER CULTURING CELLS ON PROTEIN-BLOTTED MEMBRANE
| Project/Area Number |
08670267
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Experimental pathology
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| Research Institution | NATIONAL CANCER CENTER RESEARCH INSTITUTE |
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Principal Investigator |
TERASAKI Takeo NATIONAL CANCER CENTER RESEARCH INSTITUTE,PATHOLOGY DIVISION,CHIEF RESEARCHER, 研究所・病理部, 主任研究官 (80142652)
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| Project Period (FY) |
1996 – 1997
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| Keywords | II-D cell blot method / cell bolt / protein analysis / tow-dimensional electrophoresis / small cell lung cancer / differentiation / neuronal outgrowth |
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| Research Abstract |
1.Establishment of Two-Dimensional (II-D) Cell Blot Method.
Partially digested fibronectin with trypsin was two-dimensionally electrophoresed and transferred to a membrane. Protein spots with cell binding domain on the membrane were marked after using specific antibody. Baby hamster kidney (BHK-21) cells were cultured on the membrane over night and membrane sites to where cells attached &ere marked. It was found that sites of protein spots with cell binding domain on the membrane and sites to where BHK-21 cells attached firmly were exactly the same. On the other hand, amino acid sequences of proteins transferred to the membrane can be analysed using amino acid sequencer. It was concluded that analysis of cell binding proteins could be easily done after finding a protein spots on the membrane to where cells attached and analysing amino acid sequences of the protein using amino acid sequenecer after cutting out the spot of the membrane.
2.A new protein that induced differentiation of small cell lung cancer cells to neuronal cells was determined from the conditioned medium of a lung adenocarcinoma cell line using II-D Cell Blot Method and amino acid sequencer.
3.We are planning to analyze proteins concerning cell attachment, cell differenciation or cell death using II-D cell blot method and amino acid sequencer.
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