Ras GTPase Chemi ELISA Kit

Ras GTPase Chemi ELISA Kit

Code: 52097

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.

Ras GTPase Chemi ELISA Kit Contents

The Ras GTPase Chemi ELISA Kit contains sufficient reagents to perform 96 reactions. It is shipped on dry ice and contains reagents with multiple storage temperatures inside. Please confirm receipt of all reagents upon arrival and store items at the appropriate temperatures as indicated below. All reagents are guaranteed stable for 6 months from date of receipt when stored properly. This kit includes the following components:

  • Hela whole-cell extract (EGF treated); Store at -80°C

  • GST-Raf-RBD; Store at -80°C

  • H-Ras antibody; Store at -20°C

  • Anti-rat HRP-conjugated IgG; Store at -20°C

  • Protease Inhibitor Cocktail (PIC); Store at -20°C

  • Lysis/Binding Buffer AM1; Store at 4°C

  • 10X Wash Buffer AM2; Store at 4°C

  • 10X Antibody Binding Buffer AM2; Store at 4°C

  • Chemiluminescent Reagent; Store at 4°C

  • Reaction Buffer; Store at 4°C

  • 96-well Glutathione-coated Assay Plate; Store at 4°C

  • Plate Sealer; Store at 4°C.

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.