发明名称 |
INTACT CELL ASSAY FOR PROTEIN TYROSINE PHOSPHATASES |
摘要 |
The invention relates to an intact cell assay for direct quantitation of protein tyrosine phosphatase (PTP) activity using the baculovirus expression system. A PTP expressed in transformed host insect cells is processed and localized in their predicted subcellular compartments. Assays are performed on the PTP expressing host cells challenged with a substrate such as, p- nitrophenyl phosphate. This substrate is hydrolysed to p-nitrophenol by expressed phosphatase activity. Emergence of p-nitrophenol is determined spectrophotometrically and is a measure of PTP activity. Further, the assay and transformed host cells of this invention are particularly useful in a screening method for selecting potential inhibitors to PTPs. Basically, transformed host cells are pre-incubated with or without inhibitors and the assay conducted as described. This invention relates to an intact cell assay with a direct readout of PTP activity that is useful in a cell-based screeni ng method for selecting inhibitors of PTPs.
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申请公布号 |
CA2366101(A1) |
申请公布日期 |
2000.09.14 |
申请号 |
CA20002366101 |
申请日期 |
2000.03.06 |
申请人 |
MERCK FROSST CANADA & CO. |
发明人 |
CROMLISH, WANDA A.;KENNEDY, BRIAN P. |
分类号 |
C12N1/19;C12N5/10;C12N15/09;C12N15/866;C12Q1/02;C12Q1/42;C12Q1/68;C12Q1/70;G01N33/15;G01N33/50;G01N33/68;(IPC1-7):C12Q1/42 |
主分类号 |
C12N1/19 |
代理机构 |
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地址 |
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