Ras GTPase Chemi ELISA kit 概览
Ras GTPase 背景介绍
Ras信号级联通常仅短暂激活。这是因为Ras GTPase活性随着GTP转化为GDP，蛋白质逐渐失去活性。然而，Ras基因的致癌突变可以将蛋白质锁定在活性GTP构象中，从而形成一种始终活跃的蛋白质形式。结果是组成性活跃的Ras信号和细胞生长不受控制。事实上，Ras基因突变存在于约30%的人类癌症中，尤其是胰腺癌、肺癌和结直肠癌中。因此，Ras已成为癌症研究和抗癌治疗的热门靶点。 Learn more about targeting mutant Ras proteins in our Blog: Undruggable No More - Advances for Targeting Mutant Ras Proteins and Cancer.
Ras GTPase Chemi ELISA试剂盒组分
Ras GTPase Chemi ELISA试剂盒包含足够的试剂，可完成96个反应。我们采用干冰运输，其中并包含具有多种存储温度的试剂。请在收货时确认收到所有试剂，并将物品储存在如下所示的适当温度下。所有试剂在按要求保存的情况下，可保持稳定长达6个月。该试剂盒包括以下组分：
Ras GTPase Chemi ELISA 数据展示
Ras GTPase Chemi ELISA试剂盒常见问题
Ras GTPase Chemi ELISA试剂盒引用文章
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|Ras GTPase Chemi ELISA Kit||1 x 96 rxns||52097||¥11,380||Buy|
Ras GTPase Chemi ELISA advantages
- More sensitive – assay uses only 25 µg of extract, which is 20-fold less than pull-down/Western methods
- Better results – quantitative readouts make it easier to compare results
- Less effort – no need to run gels or develop Western blots
- Save time – results in less than 5 hours
- Versatile – assay activated extracts from cells or tissue samples, or study recombinant Ras protein
Figure 1: Quantification of activated Ras in stimulated HeLa cells.
Small GTPases, also known as GTP-binding proteins, are a family of enzymes that serve as molecular switches in regulating a number of signal transduction pathways including cellular growth, apoptosis and differentiation. These switches regulate by alternating between active (GTP-bound) and inactive (GDB-bound) states. Ras GTPase is of particular interest as aberrant regulation by Ras has been implicated in a number of disease states.
The Ras GTPase Chemi ELISA method
Because activated Ras binds specifically to the Ras-binding domain (RBD) of Raf effector protein, Raf-RBD can be used as a probe to isolate Ras-GTP. The Ras GTPase Chemi ELISA Kit contains a Raf-RBD protein that is fused to GST and a 96-well, glutathione-coated assay plate. GST-Raf-RBD is first incubated on the plate for one hour to immobilize the capture probe. Addition of sample to the plate results in the binding of activated Ras to the Raf-RBD. A primary antibody specific for Ras is then added to the individual wells, followed by an HRP-conjugated secondary antibody and developing reagent. The plate is then read on a luminometer, which provides a sensitive and quantitative chemiluminescent readout of activated Ras.
Figure 1: Flow chart of the Ras GTPase Chemi ELISA Kit process.
Contents & Storage
96-well glutathione-coated assay plate with plate sealer, GST-Raf-RBD, primary antibody, HRP-conjugated secondary antibody, positive control cell extract, Protease Inhibitor Cocktail, Lysis and Reaction buffers. Reagent storage conditions vary from room temperature to -80°C, see manual for details. All reagents are guaranteed stable for 6 months when stored properly.