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    ras-gtpase-elisa-kit-chemiluminescent-ab134640.pdf

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Signal Transduction Signaling Pathway G Protein Signaling Small G Proteins Ras Family
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Ras GTPase ELISA Kit (Chemiluminescent) (ab134640)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (1)References (4)

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ELISA - Ras GTPase ELISA Kit (ab134640)

    Key features and details

    • Sensitivity: 3000 ng/well
    • Range: 3000 ng/well - 25000 ng/well
    • Sample type: Cell culture extracts, Tissue Extracts
    • Detection method: Luminescent
    • Assay type: Sandwich (quantitative)
    • Reacts with: Mouse, Human

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    Overview

    • Product name

      Ras GTPase ELISA Kit (Chemiluminescent)
    • Detection method

      Luminescent
    • Sample type

      Cell culture extracts, Tissue Extracts
    • Assay type

      Sandwich (quantitative)
    • Sensitivity

      > 3000 ng/well
    • Range

      3000 ng/well - 25000 ng/well
    • Assay time

      4h 30m
    • Assay duration

      Multiple steps standard assay
    • Species reactivity

      Reacts with: Mouse, Human
    • Product overview

      ab134640 Ras GTPase ELISA Kit is designed specifically for the study of Ras activation and can be used to study novel signaling pathways for activating Ras. The kit can also be used as a diagnostic test to detect oncogenic Ras related to malignancy. Ras GTPase ELISA Kits contain a Raf-RBD protein fused to GST that will be coated onto the provided 96-well (12 x 8 well plate), glutathione-coated plate. The activated Ras contained in cellular extract specifically binds to Raf-RBD, while inactive Ras does not bind. Bound Ras is detected by incubating with a primary antibody that detects H-Ras in mouse and H- & K-Ras in human samples. Addition of a secondary antibody conjugated to horseradish peroxidase (HRP) and developing solution provides a sensitive chemiluminescent readout that is easily quantified by luminescence. The 96-well plate with individual plate of 12 strips is suitable for manual use or high-throughput screening applications.


       


      Ras GTPase ELISA Kit specifically detects activated H- and K-Ras in Human and H-Ras in rodent samples.

    • Notes

      Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
      It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

    • Platform

      Microplate

    Properties

    • Storage instructions

      Please refer to protocols.
    • Components 1 x 96 tests
      10X Antibody Binding Buffer 1 x 2.2ml
      10X Wash Buffer 2 x 22ml
      96-well assay plate 1 unit
      Anti-rat HRP-conjugated IgG 1 x 11µl
      Chemiluminescent Reagent 1 x 2ml
      GST-Raf-RBD 4 x 25µl
      HeLa whole-cell extract (EGF treated) 2 x 40µl
      H-Ras antibody 1 x 11µl
      Lysis Buffer 1 x 50ml
      Plate sealer 1 unit
      Protease Inhibitor Cocktail 1 x 500µl
      Reaction Buffer 1 x 4ml
    • Research areas

      • Signal Transduction
      • Signaling Pathway
      • G Protein Signaling
      • Small G Proteins
      • Ras Family
      • Epigenetics and Nuclear Signaling
      • Transcription
      • Cancer susceptibility
      • Proto-oncogenes
      • Cancer
      • Signal transduction
      • G protein signaling
      • Small G proteins
      • Ras family
      • Cancer
      • Oncoproteins/suppressors
      • Oncoproteins
      • Signal transducers
      • Cancer
      • Tumor biomarkers
      • Oncoproteins
      • Kits/ Lysates/ Other
      • Kits
      • ELISA Kits
      • ELISA Kits
      • Signal transduction proteins ELISA kits
    • Function

      Ras proteins bind GDP/GTP and possess intrinsic GTPase activity.
    • Involvement in disease

      Defects in HRAS are the cause of faciocutaneoskeletal syndrome (FCSS) [MIM:218040]. A rare condition characterized by prenatally increased growth, postnatal growth deficiency, mental retardation, distinctive facial appearance, cardiovascular abnormalities (typically pulmonic stenosis, hypertrophic cardiomyopathy and/or atrial tachycardia), tumor predisposition, skin and musculoskeletal abnormalities.
      Defects in HRAS are the cause of congenital myopathy with excess of muscle spindles (CMEMS) [MIM:218040]. CMEMS is a variant of Costello syndrome.
      Defects in HRAS may be a cause of susceptibility to Hurthle cell thyroid carcinoma (HCTC) [MIM:607464]. Hurthle cell thyroid carcinoma accounts for approximately 3% of all thyroid cancers. Although they are classified as variants of follicular neoplasms, they are more often multifocal and somewhat more aggressive and are less likely to take up iodine than are other follicular neoplasms.
      Note=Mutations which change positions 12, 13 or 61 activate the potential of HRAS to transform cultured cells and are implicated in a variety of human tumors.
      Defects in HRAS are a cause of susceptibility to bladder cancer (BLC) [MIM:109800]. A malignancy originating in tissues of the urinary bladder. It often presents with multiple tumors appearing at different times and at different sites in the bladder. Most bladder cancers are transitional cell carcinomas. They begin in cells that normally make up the inner lining of the bladder. Other types of bladder cancer include squamous cell carcinoma (cancer that begins in thin, flat cells) and adenocarcinoma (cancer that begins in cells that make and release mucus and other fluids). Bladder cancer is a complex disorder with both genetic and environmental influences.
      Note=Defects in HRAS are the cause of oral squamous cell carcinoma (OSCC).
    • Sequence similarities

      Belongs to the small GTPase superfamily. Ras family.
    • Post-translational
      modifications

      Palmitoylated by the ZDHHC9-GOLGA7 complex. A continuous cycle of de- and re-palmitoylation regulates rapid exchange between plasma membrane and Golgi.
      S-nitrosylated; critical for redox regulation. Important for stimulating guanine nucleotide exchange. No structural perturbation on nitrosylation.
    • Cellular localization

      Cell membrane. Golgi apparatus membrane. The active GTP-bound form is localized most strongly to membranes than the inactive GDP-bound form (By similarity). Shuttles between the plasma membrane and the Golgi apparatus.
    • Target information above from: UniProt accession P01112 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Alternative names

      • C BAS/HAS
      • C HA RAS1
      • C-BAS/HAS
      • c-H-ras
      • C-HA-RAS1
      • CTLO
      • GTPase HRas
      • GTPase KRas
      • GTPase NRas
      • H ras
      • H RASIDX
      • H-Ras-1
      • H-RASIDX
      • Ha-Ras
      • HAMSV
      • HRAS
      • HRAS1
      • K ras
      • K RAS2A
      • K RAS2B
      • K RAS4A
      • K RAS4B
      • K-RAS
      • KRAS
      • KRAS1
      • KRAS2
      • N-RAS
      • N-terminally processed
      • NRAS
      • NRAS1
      • p21ras
      • RASH_HUMAN
      • RASH1
      • RASK2
      • Transforming protein p21
      • v Ha ras Harvey rat sarcoma viral oncogene homolog
      • v Ki ras2 Kirsten rat sarcoma viral oncogene homolog
      • v ras neuroblastoma RAS viral oncogene homolog
      see all
    • Database links

      • Entrez Gene: 4893 Human
      • Entrez Gene: 3845 Human
      • Entrez Gene: 3265 Human
      • Entrez Gene: 18176 Mouse
      • Entrez Gene: 16653 Mouse
      • Entrez Gene: 15461 Mouse
      • Omim: 190020 Human
      • Omim: 164790 Human
      • Omim: 190070 Human
      • SwissProt: P01111 Human
      • SwissProt: P01116 Human
      • SwissProt: P01112 Human
      • SwissProt: P08556 Mouse
      • SwissProt: P32883 Mouse
      • SwissProt: Q61411 Mouse
      • Unigene: 37003 Human
      • Unigene: 334313 Mouse
      see all

    Images

    • ELISA - Ras GTPase ELISA Kit (ab134640)
      ELISA - Ras GTPase ELISA Kit (ab134640)

      Quantification of activated Ras: Increasing amounts of whole-cell extracts from unstimulated 293T/17 and EGF stimulated HeLa cells were assayed for Ras activity using ab134640 Ras GTPase ELISA Kit. To illustrate the Kit’s specificity for activated Ras, 293T/17 cells which do not contain basal levels of activated Ras were used as a negative control. Data was also shown for unstimulated HeLa cells, which do contain basal levels of activated Ras. This data is provided for demonstration only.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (4)

    Publishing research using ab134640? Please let us know so that we can cite the reference in this datasheet.

    ab134640 has been referenced in 4 publications.

    • Zhou Y  et al. Puerarin improves graft bone defect through microRNA-155-3p-mediated p53/TNF-a/STAT1 signaling pathway. Int J Mol Med 46:239-251 (2020). PubMed: 32377717
    • Yin C  et al. Pharmacological Targeting of STK19 Inhibits Oncogenic NRAS-Driven Melanomagenesis. Cell 176:1113-1127.e16 (2019). PubMed: 30712867
    • Yeung Y  et al. K-Ras mutation and amplification status is predictive of resistance and high basal pAKT is predictive of sensitivity to everolimus in biliary tract cancer cell lines. Mol Oncol 11:1130-1142 (2017). PubMed: 28544747
    • Feng C  et al. miRNA-556-3p promotes human bladder cancer proliferation, migration and invasion by negatively regulating DAB2IP expression. Int J Oncol 50:2101-2112 (2017). PubMed: 28440444

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Question

    We're looking to detect RAS activation. We think your RAS GTPase ELISA kit will work well but we only have a standard ELISA plate reader for detection. The protocol states using a luminometer or a CCD camera, but we were wondering if a standard ELISA plate reader at a specific wavelength can read the results of this assay.

    Read More

    Abcam community

    Verified customer

    Asked on Apr 24 2013

    Answer


    You will need to check with the manufacturer / manual of your plate reader to make sure it is capable of reading chemiluminescence.

    Chemiluminescence is most commonly measured using a luminometer although some plate readers have an option to read chemiluminescence or can be adapted to measure total light output. If a wavelength must be selected, measurement at 425nm gives a crude indication of chemiluminescence. Some fluorescent plate readers can also be used if excitation is turned off.

    Read More

    Abcam Scientific Support

    Answered on Apr 24 2013

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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