HTRF KRAS WT GTP BIND. KIT - 500 PTS

HTRF KRAS WT GTP BIND. KIT - 500 PTS

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    产品介绍
    产品信息

    Overview

    A fast and easy way to identify new KRAS WT / GTP competitors.KRAS is a small GTPase implicated in various biological processes, such as cell proliferation, cell survival, and cell metabolism. This proto-oncogene is well known to be mutated in many cancer subtypes, inducing an uncontrolled proliferation and cell metabolism modifications. It thereby contributes to the Warburg effect in cancer cells. Like the majority of small GTPases, KRAS binds to GDP in its inactive form or binds to GTP to switch to the active form. KRAS G12C is one of the most commonly present mutant forms in cancer which lead to a permanently active state of KRAS. Identifying new KRAS / GTP competitors is therefore a relevant strategy to control biological processes involved in cancer growth by reducing the KRAS activity, as well as the associated pathways.

    Specifications

    Assay Points
    10000
    Assay Target Type
    Kit
    Assay Technology
    HTRF
    Brand
    HTRF
    Quantity
    1
    Shipping Conditions
    Shipped in Dry Ice
    Therapeutic Area
    Oncology & Inflammation
    Unit Size
    10,000 Assay Points

    Video gallery

    How it works

    Assay principle

    The HTRF KRAS GTP binding assay is in a competitive assay format, using a GTP-Red reagent as KRAS WT ligand, a 6His tagged human KRAS WT protein, and an anti 6His Cryptate-labeled antibody. The compound being tested competes with the GTP Red reagent, and thereby prevents FRET from occurring.

    1ppi-how-it-works-assay-principle-kras-wt-gtp-binding.svg

     

    Assay protocol

    The Human KRAS WT binding assay can be run in a 96- or 384-well low volume white plate (20 µL final). As described here, samples or standards are dispensed directly into the assay plate. The human His-tagged KRAS WT protein is then added, followed by the dispensing of the HTRF reagents: The anti 6His antibody labeled with Europium cryptate and the GTP-Red reagent. The reagents labeled with HTRF fluorophores may be pre-mixed and added in a single dispensing step. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally.

    2ppi-how-it-works-assay-protocol-htrf-kras-wt-gtp-binding.svg

     

    Assay details

     

    GTP-Red reagent Kd

    70nM

     

    GTP-Red reagent concentration

    50nM

     

    KRAS GTP binding Standard (GDP) IC50

    30nM

     

    KRAS GTP binding Standard Ki

     

    20.4nM

     

     

    Assay validation

    Screening of GTP competitors with human KRAS WT protein

    Various nucleotides known to be GTP competitors were added to the assay. GDP (assay standard), GTPyS, and GTP displayed the right potency, in good correlation with the literature. ATP, a non-specific nucleotide, and BAY-293, a KRAS/SOS1 inhibitor, had no effect on the assay, confirming the specificity of the kit.

    3assay-validation-human-kras-wt-gtp-binding-1.svg
    4assay-validation-human-kras-wt-gtp-binding-2.svg

     

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    货号:
    64BDKRASWPEG
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    询价
    500Tests
    10000Tests
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