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| Prokaryotic Expression of LINE-1 Encoded Protein L1-ORF1 and Preparation of Anti-L1-ORF1 Antibody |
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Abstract [Abstract] AIM: To detect L1-ORF1, an ubiquitous protein in tumor tissue, we prepare its polyclonal antibody with rabbits. METHODS: L1-ORF1 gene was prepared by PCR amplification and cloned into pGEX-6P-1 with GST fusion. After expressing in E.coli, GST-L1-ORF1 fusion protein was purified by Glutathione Sepharose 4B and subjected to immunity with rabbits. The titer of anti-serum was measured by ELISA and the specificity was examined by Western Blot with the cell lysates derived from HepG2 tumor cells. RESULT: The anti-L1-ORF1p antibody exhibits a high sensitivity and specificity, which was evaluated by L1-ORF1p transfected normal cells and tumor cells. CONCLUSION：We successfully acquired anti L1-ORF1p polyclonal antibody with high sensitivity and specificity.
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Received: 07 June 2010
Published: 19 October 2010
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