RECEPTOR TYROSINE KINASE ASSAYS
    2.
    发明申请
    RECEPTOR TYROSINE KINASE ASSAYS 审中-公开
    受体酪氨酸激酶测定

    公开(公告)号:US20120040372A1

    公开(公告)日:2012-02-16

    申请号:US13280919

    申请日:2011-10-25

    IPC分类号: G01N33/573

    CPC分类号: G01N33/566 C12Q1/34 C12Q1/485

    摘要: Methods for detecting phosphorylation of receptor tyrosine kinases (“RTKs”) upon activation and the modulation of activation by a candidate compound are provided. The method employs cells comprising two fusion products: (1) an RTK fused to a small fragment of β-galactosidase and (2) a phosphotyrosine binding peptide fused to the large fragment of β-galactosidase, where the 2 fragments weakly complex to form an active enzyme, and optionally a construct for a cytosolic RTK phosphorylating kinase, when the RTK does not autophosphoryate. To detect phosphorylation a β-galactosidase substrate is added to the cells, whereby product formation indicates the occurrence of phosphorylation.

    摘要翻译: 提供了激活后受体酪氨酸激酶(“RTK”)磷酸化和候选化合物激活调节的方法。 该方法使用包含两个融合产物的细胞:(1)融合到β-半乳糖苷酶的小片段的RTK和(2)与β-半乳糖苷酶的大片段融合的磷酸酪氨酸结合肽,其中2个片段与 形成活性酶,以及任选的胞质RTK磷酸化激酶的构建体,当RTK不是自磷酸化时。 为了检测磷酸化,将一个半乳糖苷酶底物加入到细胞中,由此产物形成指示磷酸化的发生。

    Receptor Tyrosine Kinase Assays
    3.
    发明申请
    Receptor Tyrosine Kinase Assays 有权
    受体酪氨酸激酶测定

    公开(公告)号:US20100041052A1

    公开(公告)日:2010-02-18

    申请号:US12536667

    申请日:2009-08-06

    IPC分类号: C12Q1/68 G01N33/573

    CPC分类号: G01N33/566 C12Q1/34 C12Q1/485

    摘要: Methods for detecting phosphorylation of receptor tyrosine kinases (“RTKs”) upon activation are provided. The method employs cells comprising two fusion products: (1) an RTK fused to a small fragment of β-galactosidase and (2) a phosphotyrosine binding peptide fused to the large fragment of β-galactosidase, where the 2 fragments weakly complex to form an active enzyme, and optionally a construct for a cytosolic RTK phosphorylating kinase, when the RTK does not autophosphoryate. To detect phosphorylation a β-galactosidase substrate is added to the cells, whereby product formation indicates the occurrence of phosphorylation.

    摘要翻译: 提供了激活后检测受体酪氨酸激酶(“RTK”)磷酸化的方法。 该方法使用包含两个融合产物的细胞:(1)融合到β-半乳糖苷酶的小片段的RTK和(2)与β-半乳糖苷酶的大片段融合的磷酸酪氨酸结合肽,其中2个片段与 形成活性酶,以及任选的胞质RTK磷酸化激酶的构建体,当RTK不是自磷酸化时。 为了检测磷酸化,将一个半乳糖苷酶底物加入到细胞中,由此产物形成指示磷酸化的发生。

    Receptor tyrosine kinase assays
    4.
    发明授权
    Receptor tyrosine kinase assays 有权
    受体酪氨酸激酶测定

    公开(公告)号:US08067155B2

    公开(公告)日:2011-11-29

    申请号:US12536667

    申请日:2009-08-06

    IPC分类号: C12Q1/00 G01N33/53

    CPC分类号: G01N33/566 C12Q1/34 C12Q1/485

    摘要: Methods for detecting phosphorylation of receptor tyrosine kinases (“RTKs”) upon activation are provided. The method employs cells comprising two fusion products: (1) an RTK fused to a small fragment of β-galactosidase and (2) a phosphotyrosine binding peptide fused to the large fragment of β-galactosidase, where the 2 fragments weakly complex to form an active enzyme, and optionally a construct for a cytosolic RTK phosphorylating kinase, when the RTK does not autophosphoryate. To detect phosphorylation a β-galactosidase substrate is added to the cells, whereby product formation indicates the occurrence of phosphorylation.

    摘要翻译: 提供了激活后检测受体酪氨酸激酶(“RTK”)磷酸化的方法。 该方法使用包含两个融合产物的细胞:(1)融合到β-半乳糖苷酶的小片段的RTK和(2)与β-半乳糖苷酶的大片段融合的磷酸酪氨酸结合肽,其中2个片段与 形成活性酶,以及任选的胞质RTK磷酸化激酶的构建体,当RTK不是自磷酸化时。 为了检测磷酸化,将一个半乳糖苷酶底物加入到细胞中,由此产物形成指示磷酸化的发生。

    Detection of protein translocation by beta-galactosidase reporter fragment complementation
    5.
    发明申请
    Detection of protein translocation by beta-galactosidase reporter fragment complementation 有权
    通过β-半乳糖苷酶报道片段互补检测蛋白质易位

    公开(公告)号:US20050287522A1

    公开(公告)日:2005-12-29

    申请号:US11132764

    申请日:2005-05-18

    摘要: Methods and compositions are provided for detecting molecular translocations, particularly protein translocations within and between subcellular copartments, using at least two components that exhibit a localization-dependent difference in complementation activity. In particular, alpha-complementing β-galactosidase fragments are provided. These β-galactosidase reporter fragments display significantly enhanced enzymatic activity when one fragment is localized in a membrane. Methods for carrying out no-wash ELISA assays based on the reporter component system are also provided.

    摘要翻译: 提供了用于检测分子易位,特别是在亚细胞间室内和之间的蛋白质易位的方法和组合物,其使用至少两种显示互补活性的定位依赖性差异的组分。 特别地,提供了α-补体的β-半乳糖苷酶片段。 当一个片段定位在膜中时,这些β-半乳糖苷酶报告物片段显示出显着增强的酶活性。 还提供了基于报道分子系统进行免洗ELISA测定的方法。