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
17-497
96 assays  
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      Overview

      Key Spec Table

      Analytes AvailableSpecies ReactivityKey Applications
      RasR, H, MCell Function Assay, ELISA
      Description
      Catalogue Number 17-497
      Brand Family Upstate
      Trade Name
      • Upstate
      Description Ras Activation ELISA ASSAY Kit
      Background Information 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.
      References
      Product Information
      Components 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.
      Applications
      Key Applications
      • Cell Function Assay
      • ELISA
      Biological Information
      Species Reactivity Rat Human Mouse
      Analytes Available
      • Ras
      Entrez Gene Number
      Entrez Gene Summary 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]
      Gene Symbol
      • 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
      UniProt Number
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality Assurance Routinely evaluated by ELISA.
      Usage Statement
      • 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.
      Storage and Shipping Information
      Packaging Information
      Material Size 96 assays
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      MSDS

      Languages
      English

      Certificates of Analysis

      TitleLot Number
      Ras Activation ELISA ASSAY Kit 2477960
      Ras Activation ELISA ASSAY Kit - 2137027 2137027
      Ras Activation ELISA ASSAY Kit - 2330563 2330563
      Ras Activation ELISA ASSAY Kit - 2381828 2381828
      Ras Activation ELISA ASSAY Kit - 2382602 2382602
      Ras Activation ELISA ASSAY Kit - 2393894 2393894
      Ras Activation ELISA ASSAY Kit - 2445108 2445108
      Ras Activation ELISA ASSAY Kit - 1969065 1969065
      Ras Activation ELISA ASSAY Kit - 1975943 1975943
      Ras Activation ELISA ASSAY Kit - 1982495 1982495
      Ras Activation ELISA ASSAY Kit - 1997007 1997007
      Ras Activation ELISA ASSAY Kit - 2035086 2035086
      Ras Activation ELISA ASSAY Kit - 2198971 2198971
      Ras Activation ELISA ASSAY Kit - 2203064 2203064
      Ras Activation ELISA ASSAY Kit - 2287713 2287713
      Ras Activation ELISA ASSAY Kit - 2518473 2518473
      Ras Activation ELISA ASSAY Kit - 2528702 2528702
      Ras Activation ELISA ASSAY Kit - JBC1771117 JBC1771117
      Ras Activation ELISA ASSAY Kit - NG1633510 NG1633510
      Ras Activation ELISA ASSAY Kit - NG1667741 NG1667741
      Ras Activation ELISA ASSAY Kit - NG1749753 NG1749753
      Ras Activation ELISA ASSAY Kit - NG1829837 NG1829837
      Ras Activation ELISA ASSAY Kit - NG1833929 NG1833929
      Ras Activation ELISA ASSAY Kit - NG1851604 NG1851604
      Ras Activation ELISA ASSAY Kit - NG1859587 NG1859587
      Ras Activation ELISA ASSAY Kit - NG1898614 NG1898614
      Ras Activation ELISA ASSAY Kit - NG1922973 NG1922973
      Ras Activation ELISA ASSAY Kit - NG1941601 NG1941601
      Ras Activation ELISA ASSAY Kit - RE0810064 RE0810064
      Ras Activation ELISA ASSAY Kit -2510248 2510248

      Brochure

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

      User Guides

      Title
      Instruction Manual For Ras Activation ELISA ASSAY Kit

      Product Insert

      Title
      Instruction Manual For Ras Activation ELISA ASSAY Kit

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      Categories

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