• 我要登录|
  • 免费注册
    |
  • 我的丁香通
    • 企业机构:
    • 成为企业机构
    • 个人用户:
    • 个人中心
  • 移动端
    移动端
丁香通 logo丁香实验_LOGO
搜实验

    大家都在搜

      大家都在搜

        0 人通过求购买到了急需的产品
        免费发布求购
        发布求购
        点赞
        收藏
        wx-share
        分享

        Assaying AKT/Protein Kinase B Activity

        互联网

        485
        Akt/protein kinase B (PKB) is a serine/threonine kinase that mediates many of the anabolic actions of insulin ( 1 ), as well as the growth-promoting and/or antiapoptotic effects of other growth factors, cytokines, or transforming oncogenes ( 2 ). These agonists all stimulate Akt/PKB by promoting its phosphorylation on two regulatory residues (e.g., S473 and T308 for the Akt1 isoform), an event dependent on the prior activation of a signaling pathway initiated by the lipid kinase phosphatidylinositol 3-kinase. Once Akt/PKB is activated it phosphorylates numerous different substrates, including transcription factors (e.g., FKHR1, others), anti-apoptotic enzymes (e.g., Bad caspase-9), and metabolic enzymes (e.g., glycogen synthase kinase 3β), to regulate this diverse array of biological processes. Herein we describe a method for measuring the catalytic activity of Akt/PKB isolated from cell or tissue extracts by quantifying its ability to catalyze phosphate incorporation into an exogenous substrate (outlined in Fig. 1 ). This chapter illustrates techniques for immunoprecipitating Akt/PKB, choosing an appropriate substrate for the reaction, and optimizing the assay conditions.
        Fig. 1.  Schematic diagram depicting the major steps in the Akt/PKB kinase assay.

        ad image
        提问
        扫一扫
        丁香实验小程序二维码
        实验小助手
        丁香实验公众号二维码
        扫码领资料
        反馈
        TOP
        打开小程序