Ras G-LISA Activation Assay Kit (Colorimetric Based) - 96 assays

G-LISA Ras Activation Assay Biochem Kit (Colorimetric format)
$0.00

Product Uses Include

  • Ras signaling pathway studies
  • Ras activation assays with primary cells
  • Studies of Ras activators and inactivators
  • Ras activation assays with limited material
  • High throughput screens for Ras activation

Introduction

With the new Ras G-LISA™ kit you can now measure Ras activation from cell and tissue samples in less than 3 h.  G-LISA™ requires only 1-5% of the material needed for a conventional pull-down assay.  You will also be able to handle large sample numbers and generate quantitative results.  For a more detailed introduction on G-LISA™ assays and a listing of other available G-LISA™ kits, see our main G-LISA™ page.

The Ras G-LISA™ kit contains a Ras GTP-binding protein linked to the wells of a 96 well plate. Active, GTP-bound Ras in cell/tissue lysates will bind to the wells while inactive GDP-bound Ras is removed during washing steps.  The bound active Ras is detected with a Ras specific antibody.  The degree of Ras activation is determined by comparing readings from activated cell lysates versus non-activated cell lysates.  Inactivation of Ras is generally achieved in tissue culture by a serum starvation step (see kit datasheet for more information).  

Kit contents

The kit contains sufficient reagents to perform 96 Ras activation assays. Since the Ras-GTP affinity wells are supplied as strips and the strips can be broken into smaller pieces, each kit can be used for anywhere from one to multiple assays. The following components are included in the kit: 

  1. 96 Ras-GTP affinity wells (divisible into 12 strips of 8 wells each)
  2. Lysis buffer
  3. Binding buffer
  4. Antigen presenting buffer
  5. Wash buffer
  6. Antibody dilution buffer
  7. Anti-Ras antibody
  8. HRP-labeled secondary antibody
  9. Positive control Ras protein
  10. Protease inhibitor cocktail (Cat. # PIC02)
  11. Absorbance detection reagents
  12. Precision Red™ Advanced protein assay reagent (Cat. # ADV02)
  13. Manual with detailed protocols and extensive troubleshooting guide

    Equipment needed

  14. 96-well plate spectrophotometer capable of reading 490 nm wavelength
  15. Multichannel or multidispensing pipettor
  16. Orbital microplate shaker capable of at least 200 rpm shaking (400 rpm is optimal)

Example results

BK131-Fig1

Figure 1.  Ras  activation by EGF measured by G-LISA™.  HeLa cells were serum starved (SS) for 24 h and treated with EGF (100 ng/ml for 2 min). 25,  12.5, 5, 1.25 µg of cell lysates were subjected to the G-LISA™ assay. Absorbance was read at 490 nm.  Data are background subtracted

Go to main G-LISA™ page

G-LISA Products:
Cdc42 G-LISA™ Activation Assay, colorimetric format (Cat.# BK127)
Rac1 G-LISA™ Activation Assay, luminescence format (Cat.# BK126)
G-LISA Rac 1,2,3 Activation Assay Biochem Kit (colorimetric format (Cat.# BK125)
RhoA G-LISA™ Activation Assay, colorimetric format (Cat.# BK124)
RhoA G-LISA™ Activation Assay, luminescence format (Cat.# BK121)

Associated Products:
Anti-Cdc42 monoclonal antibody (Cat.# ACD03)
Anti-Rac1 monoclonal antibody (Cat.# ARC03)
Anti-RhoA monoclonal antibody (Cat.# ARH03)

For product Datasheets and MSDSs please click on the PDF links below.   

 

    AuthorTitleJournalYearArticle Link
    Nataraj, Nishanth Belugali et al.Nucleoporin-93 reveals a common feature of aggressive breast cancers: robust nucleocytoplasmic transport of transcription factorsCell Reports2022ISSN 2211-1247
    Swaminathan, Bhairavi et al.Endothelial Notch signaling directly regulates the small GTPase RND1 to facilitate Notch suppression of endothelial migrationScientific Reports2022ISSN 2045-2322
    Hofmann, Marco H. et al.Bi-3406, a potent and selective sos1–kras interaction inhibitor, is effective in kras-driven cancers through combined mek inhibitionCancer Discovery2021ISSN 2159-8290
    Mao, De Yu et al.CLIC1 and CLIC4 mediate endothelial S1P receptor signaling to facilitate Rac1 and RhoA activity and functionScience Signaling2021ISSN 1937-9145
    Tisi, Renata et al.The Multi-Level Mechanism of Action of a Pan-Ras Inhibitor Explains its Antiproliferative Activity on Cetux****-Resistant Cancer CellsFrontiers in Molecular Biosciences2021ISSN 2296-889X
    Lu, Hengyu et al.Research paper resistance to allosteric SHP2 inhibition in FGFR-driven cancers through rapid feedback activation of FGFROncotarget2020ISSN 1949-2553
    Wang, Xuesong et al.Sevof****** inhibits growth factor-induced angiogenesis through suppressing Rac1/paxillin/FAK and Ras/Akt/mTORFuture oncology (London, England)2020ISSN 1744--8301
    Kim, Jin Hee et al.Rational design of small molecule RHOA inhibitors for gastric cancerThe Pharmacogenomics Journal 2020 20:42020ISSN 1473--1150
    Morgan, Chase J. et al.Endogenous IQGAP1 and IQGAP3 do not functionally interact with RasScientific Reports2019ISSN 2045-2322
    Casique-Aguirre, Diana et al.KRas4B-PDE6δ complex stabilization by small molecules obtained by virtual screening affects Ras signaling in pancreatic cancer 06 Biological Sciences 0601 Biochemistry and Cell BiologyBMC Cancer2018ISSN 1471-2407
    Zhao, Tuo et al.Simulated Microgravity Reduces Focal Adhesions and Alters Cytoskeleton and Nuclear Positioning Leading to Enhanced Apoptosis via Suppressing FAK/RhoA-Mediated mTORC1/NF-κB and ERK1/2 PathwaysInternational Journal of Molecular Sciences2018ISSN 1422-0067
    Li, Qing fen et al.Activation of Ras in the Vascular Endothelium Induces Brain Vascular Malformations and Hemorrhagic StrokeCell Reports2018ISSN 2211-1247
    Wong, Gabrielle S. et al.Targeting wild-type KRAS-amplified gastroesophageal cancer through combined MEK and SHP2 inhibition letterNature Medicine2018ISSN 1546-170X
    Huang, Jia Lin et al.Lipoprotein-biomimetic nanostructure enables efficient targeting delivery of siRNA to Ras-activated glioblastoma cells via macropinocytosisNature Communications2017ISSN 2041-1723
    Chen, Guo Ping et al.Inhibition of farnesyl pyrophosphate synthase attenuates high glucose-induced vascular smooth muscle cells proliferationMolecular Medicine Reports2017ISSN 1791-3004
    Zhao, Chen Ze et al.Inhibition of farnesyl pyrophosphate synthase improves pressure overload induced chronic cardiac remodelingScientific Reports2016ISSN 2045-2322
    Chen, Bin et al.Alteration of mevalonate pathway related enzyme expressions in pressure overload-induced cardiac hypertrophy and associated heart failure with preserved ejection fractionCellular Physiology and Biochemistry2013ISSN 1421-9778
    Camargo, Livia De Lucca et al.Endo-PDI is required for TNFα-induced angiogenesisFree Radical Biology and Medicine2013
    Lee, Sei Jung et al.Regulation of Hypoxia-inducible Factor 1α (HIF-1α) by Lysophosphatidic Acid Is Dependent on Interplay between p53 and Krüppel-like Factor 5The Journal of Biological Chemistry2013ISSN 0021-9258
    Gil-Henn, H. et al.Arg/Abl2 promotes invasion and attenuates proliferation of breast cancer in vivoOncogene2013ISSN 0950-9232

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