17-497 | Ras Activation ELISA ASSAY Kit

Ras Activation ELISA Assay Kit Performance
Following the protocol outlined in the manual, 50 g of EGF-stimulated HeLa cell lysate (50 ng/mL EGF for 1 minute) shows elevated levels of activated Ras compared with the basal levels of activated Ras in 50 μg of unstimulated HeLa cell lysate. Assay background, which contains no cell lysate, is included to show the signal over noise ratio.
Insert: Western blot of Ras pull-down using 10g Raf1-RBD (14-278). Lysate was prepared from HeLa cells serum starved overnight, plus or minus 1 minute stimulation with 50 ng/mL EGF.
Ras Activation ELISA Assay Kit Performance
Following the protocol outlined in the manual, 50 g
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      Overview

      Replacement Information

      Tabla espec. clave

      Analitos disponibles Reactividad según especies Aplicaciones clave
      RasRata, Ser humano, RatónELISA
      Descripción
      Número de catálogo 17-497
      Replaces 17-424
      Marca de familia Upstate
      Nombre comercial
      • Upstate
      Descripción Ras Activation ELISA ASSAY Kit
      Información preliminar Ras proteins function as GDP/GTP-regulated binary switches in signal transduction cascades that can lead to cell growth, proliferation, differentiation, or survival. This Ras superfamily has at least five major branches that include Ras, Rho, Ran, Arf/Sar, Rab. Ras itself has 3 primary isoforms (H-Ras, N-Ras, and K-Ras) that differ in their approximately 20 C-terminal amino acids. They are collectively referred to as Ras. In its active form, Ras is bound to GTP. This causes a conformational change that allow it to interact and bind to several effector molecules, most notably the members of the Raf family, the RalGDS family, and Phosphoinositide 3-kinases (PI3 Kinase). Ras then cleaves GTP to GDP resulting in its inactivation. In its oncogenic, mutated state, Ras is unable to hydrolyze GTP to GDP, thus staying in an active state and activating numerous pathways.
      Due to its role as a key regulator of cellular functions and its implications in various cancers, Ras has been a popular target for cancer research and anti-cancer therapeutics for the past two decades for both academic and pharmaceutical research. It was also the first human oncogene identified. Mutations in cellular Ras have been found to be present in a large percentage of all human cancers. More specifically, K-Ras mutations occur frequently in lung, pancreatic, and colon cancers, where as H-Ras mutations are prevalent in bladder, kidney, and thyroid cancers, and N-Ras mutations are associated with melanoma, hepatocellular carcinoma, and leukemia (3). In recent studies, Ras activity has also been implicated in a number of human development defects such as Costello syndrome and Noonan syndrome (4).
      One path that the pharmaceutical industry has taken to control Ras and its activity is by attacking what some consider its Achilles’ heel. For its activation, Ras must localize to the plasma membrane, but interestingly, it lacks a transmembrane domain. To achieve this, Ras must first undergo a post-translational modification known as prenylation or geranylation at its C-terminal CAAX motif. These drugs have yet to pass clinical trials though and there is doubt that they will ever be successful in treating tumors associated with Ras activation.
      Referencias bibliográficas
      Información del producto
      Componentes
      • 11. EGF stimulated HeLa Cell Lysate: (Part No. 12-500) 2 vials containing 40 µL of lysate at 5 mg/mL concentration, lysed in Mg2+ Lysis/Wash buffer (Catalog # CS202617).
      • 1. Glutathione-Coated Plate, 96-well strip format: (Part No. 3007063) 1 plate containing 12 x 8 strip-wells coated with glutathione.
      • 2. Anti-Ras Detection Antibody: (Part No. 2006992) One vial containing 10 μL of anti-Ras mouse monoclonal primary antibody. Can specifically detect K-, H-, N- isoforms of Ras.
      • 3. Mg2+ Lysis/Wash Buffer, 5X: (Part No. CS202617) 1 vial containing 18 mL of 5X Mg2+ Lysis/Wash Buffer for preparation of cell lysate.
      • 4. 20% Tween® 20: (Part No. # 20-246) One vial containing 3 mL of 20% Tween® 20.
      • 5. 10% BSA in TBS (Blocking Buffer): (Part No. 20-191) One bottle containing 10 mL 10% BSA in 1X TBS.
      • 6. 20X TBS Wash: (Part No. 20-190) One bottle containing 50 mL of 20X TBS, pH 7.4.
      • 7. Gt x Ms, HRP: (Part No. CS202619) One amber vial containing 25 µL of goat anti-mouse HRP conjugated antibody.
      • 8. Chemiluminescent Reaction Buffer: (Part No. 90496) One amber bottle containing 8 mL of Chemiluminescent Reaction Buffer. Keep out of light.
      • 9. Chemiluminescent Detection Reagent: (Part No. 90495) One bottle vial containing 4 mL of Chemiluminescent Detection Reagent. Keep out of light.
      • Store at -80º C
      • 10. Raf-1-RBD, GST: (Part No. 14-863) 2 vials containing 150 μL of Raf-1-RBD at 2 μg/μL.
      Aplicaciones
      Aplicaciones clave
      • ELISA
      Información biológica
      Reactividad según especies Rata Ser humano Ratón
      Analitos disponibles
      • Ras
      Número de Entrez Gene
      Resumen de Entrez Gene K-Ras
      This gene, a Kirsten ras oncogene homolog from the mammalian ras gene family, encodes a protein that is a member of the small GTPase superfamily. A single amino acid substitution is responsible for an activating mutation. The transforming protein that results is implicated in various malignancies, including lung adenocarcinoma, mucinous adenoma, ductal carcinoma of the pancreas and colorectal carcinoma. Alternative splicing leads to variants encoding two isoforms that differ in the C-terminal region. [provided by RefSeq]
      H-Ras
      This gene belongs to the Ras oncogene family, whose members are related to the transforming genes of mammalian sarcoma retroviruses. The products encoded by these genes function in signal transduction pathways. These proteins can bind GTP and GDP, and they have intrinsic GTPase activity. This protein undergoes a continuous cycle of de- and re-palmitoylation, which regulates its rapid exchange between the plasma membrane and the Golgi apparatus. Mutations in this gene cause Costello syndrome, a disease characterized by increased growth at the prenatal stage, growth deficiency at the postnatal stage, predisposition to tumor formation, mental retardation, skin and musculoskeletal abnormalities, distinctive facial appearance and cardiovascular abnormalities. Defects in this gene are implicated in a variety of cancers, including bladder cancer, follicular thyroid cancer, and oral squamous cell carcinoma. Multiple transcript variants, which encode different isoforms, have been identified for this gene. [provided by RefSeq]
      Símbolo de gen
      • neuroblastoma RAS viral (v-ras) oncogene homolog
      • KRAS
      • K-Ras
      • v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog
      • K-ras p21 protein
      • HRAS
      • H-Ras
      • Ha-Ras
      • NRAS
      • N-Ras
      Número UniProt
      Información fisicoquímica
      Dimensiones
      Información de materiales
      Información toxicológica
      Información de seguridad según el GHS
      Información de seguridad
      Declaraciones de uso del producto
      Aseguramiento de calidad Routinely evaluated by ELISA.
      Declaración de uso
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Información de almacenamiento y transporte
      Información sobre embalaje
      Cantitad 96 assays
      Información de transporte
      Información complementaria
      Especificaciones

      Documentation

      Ficha datos de seguridad (MSDS)

      Idiomas
      English

      Certificados de análisis

      CargoNúmero de lote
      Ras Activation ELISA ASSAY Kit2477960
      Ras Activation ELISA ASSAY Kit - 21370272137027
      Ras Activation ELISA ASSAY Kit - 23305632330563
      Ras Activation ELISA ASSAY Kit - 23818282381828
      Ras Activation ELISA ASSAY Kit - 23826022382602
      Ras Activation ELISA ASSAY Kit - 23938942393894
      Ras Activation ELISA ASSAY Kit - 24451082445108
      Ras Activation ELISA ASSAY Kit - 19690651969065
      Ras Activation ELISA ASSAY Kit - 19759431975943
      Ras Activation ELISA ASSAY Kit - 19824951982495
      Ras Activation ELISA ASSAY Kit - 19970071997007
      Ras Activation ELISA ASSAY Kit - 20350862035086
      Ras Activation ELISA ASSAY Kit - 21989712198971
      Ras Activation ELISA ASSAY Kit - 22030642203064
      Ras Activation ELISA ASSAY Kit - 22877132287713
      Ras Activation ELISA ASSAY Kit - 25184732518473
      Ras Activation ELISA ASSAY Kit - 25287022528702
      Ras Activation ELISA ASSAY Kit - JBC1771117JBC1771117
      Ras Activation ELISA ASSAY Kit - NG1633510NG1633510
      Ras Activation ELISA ASSAY Kit - NG1667741NG1667741
      Ras Activation ELISA ASSAY Kit - NG1749753NG1749753
      Ras Activation ELISA ASSAY Kit - NG1829837NG1829837
      Ras Activation ELISA ASSAY Kit - NG1833929NG1833929
      Ras Activation ELISA ASSAY Kit - NG1851604NG1851604
      Ras Activation ELISA ASSAY Kit - NG1859587NG1859587
      Ras Activation ELISA ASSAY Kit - NG1898614NG1898614
      Ras Activation ELISA ASSAY Kit - NG1922973NG1922973
      Ras Activation ELISA ASSAY Kit - NG1941601NG1941601
      Ras Activation ELISA ASSAY Kit - RE0810064RE0810064
      Ras Activation ELISA ASSAY Kit -25102482510248
      Ras Activation ELISA ASSAY Kit -25746082574608

      Folleto

      Cargo
      Product Selection Guide - Antibodies, small molecule inhibitors, kits, assays and proteins for signaling research.

      Manuales del usuario

      Cargo
      Instruction Manual For Ras Activation ELISA ASSAY Kit

      Prospecto del producto

      Cargo
      Instruction Manual For Ras Activation ELISA ASSAY Kit

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      Categorías

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      Life Science Research > Antibodies and Assays > Immunoassays > Enzyme-linked Immunosorbent Assay (ELISA) > Complete ELISA Kits