Ras GTPase Chemi ELISA Kit

Synonyms

Ras GTPase Chemi ELISA Kit, 52097, ras, gtpase, ras gtpase, small gtpase, gtp, elisa, chemiluminescent

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Overview

Ras GTPase Chemi ELISA Kit Overview

The Ras GTPase Chemi ELISA Kit was designed specifically for the study of Ras activation, and can be used to detect and quantify activated Ras GTPase in cell or tissue extracts. Monitoring levels of activated Ras can be useful for studying novel Ras activating signaling pathways, detecting oncogenic Ras proteins, or monitoring the effects of possible therapeutic compounds.

The Ras GTPase Chemi ELISA Kit uses the unique binding properties of the Ras-binding domain of the c-Raf kinase protein (Raf-RBD) which only binds activated Ras. A GST-fused Raf-RBD protein is bound to a 96-well glutathione-coated assay plate and 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 chemiluminescent readout of activated Ras. The 96-well plate is suitable for both manually assaying small numbers of samples and for use in automated high-throughput screening assays.

Flow Chart of Ras GTPase Chemi ELISA Kit
(Click image to enlarge)

Ras GTPase Background

The Ras protein is a GTPase enzyme. GTPases are a family of enzymes which bind to and hydrolyze GTP, functioning as molecular switches. When bound to GTP, Ras is active and activates effector proteins involved in the downstream control of cell proliferation and differentiation. When bound to GDP, Ras is inactive.

Ras signaling cascades are normally only transiently activated. This is because the Ras GTPase activity gradually converts GTP to GDP, rendering the protein inactive. However, oncogenic mutations of the Ras gene can lock the protein into the active GTP conformation, creating a form of the protein that is always active. The result is constituatively active Ras signaling and cells which grow uncontrollably. In fact, mutations of the Ras gene are present in around 30% of human cancers and are present in an especially high percentage of pancreatic, lung, and colorectal cancers. Consequently, Ras has become a popular target for cancer research and anti-cancer therapeutics. 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 Kit Highlights

• Detection Range – Works with 3-25 µg whole cell extract or > 0.6 ng purified protein per well.
• Better Results – ELISA format is more sensitive than pull-down/Western methods.
• Versatile – Use with extracts from cell and tissue samples, or with purified Ras protein
• Reactivity – detects activated human K-Ras and H-Ras or activated rodent H-Ras
• Quick – Results in fewer than 5 hours