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®<strong>Life</strong> <strong>Sciences</strong>Therapeutics®WNT PATHWAYIntegrated solutionsfor screening Wnt regulatorsClinical LabsImage: TR Arnett, UCL
SOLUTIONS FORWNT PATHWAY SCREENINGHigh-throughput Assays for Canonical Wnt Pathway AnalysisSince the discovery of the Wnt gene over 30 years ago <strong>as</strong> the Int1 proto-oncogene involved inthe pathology of Mouse Mammary Tumor Virus (MMTV), Wnt expression and signal transductionhave been identified <strong>as</strong> critical components in regulating embryogenesis, cell proliferation,and bone, glucose, and lipid metabolism. Perturbations in Wnt expression, surface receptors,signaling partners (including β-catenin and GSK-3β), and the battery of Wnt-regulated genes(collectively the canonical Wnt pathway) are known to manifest in a variety of clinicallyrelevant dise<strong>as</strong>e states, including cancer, osteoporosis, cardiov<strong>as</strong>cular dise<strong>as</strong>e, diabetes, andneurodegeneration.WNT MODULATOR SCREENINGWntWIF-1PATHWAY ANALYSISScreen-Well ®Wnt PathwayLibrarySOSTDKKLRP5/6KremenLRP5/6WntFrizzledsFRPProte<strong>as</strong>omaldegradationLRP5/6WntFrizzledAxin Frat-1DshELISAsDkk-1, GSK-3β,[pSer9] GSK-3β,β-cateninLeading Light TMWnt ReporterAssayAPCGSK3AxinPβ-CateninGSK3β-CateninNuclearLocalizationp300/CBPβ-Cateninβ-Cateninβ-CateninWnt Regulated Gene ExpressionTCF/LEFAntibodiesGSK-3β,GSK-3 α/β,Dishevelled-1,Dishevelled-2,β-cateninProteinsAPC, Dkk-1,GSK-3β, Wnt-3a,C<strong>as</strong>ein Kin<strong>as</strong>e IIWnt InhibitionReceptor Activationβ-catenin phosphorylation by GSK3ββ-catenin degredationWnt ActivationReceptor Activationβ-catenin phosphorylation by GSK3ββ-catenin nuclear localizationCellular AnalysisWe have extensive experience in buildingnext generation cell b<strong>as</strong>ed <strong>as</strong>says for use indrug development, secondary screening, andpathway analysis. Our Leading Light andCELLestial ® <strong>as</strong>say systems are optimizedto meet the needs of demanding highthroughputapplications to deliver better,high-quality hits.Our portfolio of over 300 ELISA kits includessensitive, specific <strong>as</strong>says for key signalingproteins in the canonical Wnt pathway in avariety of species. In addition, our catalogof over 3000 antibodies and recombinantproteins includes thoroughly validatedreagents for dissection of Wnt pathwaysignaling.Small Molecule ChemistryOur long-standing, flagship Screen-Well ®Compound Library portfolio offers ane<strong>as</strong>y, ready-to-use method to streamlinecompound screening. Included within ourthousands of stand-alone small molecules isa unique collection of Wnt Pathway targetingcompounds <strong>as</strong> well <strong>as</strong> focused libraries forkin<strong>as</strong>e inhibitors, natural products, prote<strong>as</strong>einhibitors, nuclear receptors, autophagyregulators, and more.Through our supplier network built over a 25year history, we have the ability to rapidlyand inexpensively source traditional, custom,and bulk compounds in the gram to kilogramscale. We complete our sourcing usingstringent quality control standards withstate-of-the-art methods.
SCREEN WNT PATHWAY ACTIVATORS & INHIBITORSDevelopment of Wnt pathway modulating drugs requires the ability to rapidly screen the effect of novel or repurposed compounds andbiological therapeutics on the expression of Wnt-regulated genes, and confirmation of such effects by related changes in key signalingcomponents. Together, the Leading Light Wnt Reporter Assay and Screen-Well® Wnt Pathway Library facilitate high-throughputanalysis of Wnt-related signaling.Leading Light Wnt Reporter Assay (ENZ-61001)Reliable, All-inclusive Screening of Wnt Pathway ModulatorsThe Leading Light Wnt Reporter Assay Starter Kit is a high-sensitivity, lucifer<strong>as</strong>e-b<strong>as</strong>ed <strong>as</strong>say for the analysis of canonicalWnt signaling. The <strong>as</strong>say uses a 3T3 mouse fibrobl<strong>as</strong>t cell line modified with a lucifer<strong>as</strong>e reporter gene under the control ofWnt-responsive promoters (TCF/LEF). The chemiluminescent readout is amenable to both 96-well and 384-well plate formats,facilitating high-throughput analysis of Wnt pathway activators and inhibitors.Screen your Wnt Pathwayin only 4 steps!All kit components included!BENEFITS• A true end-point detection system for screening compounds• Greater sensitivity than other whole cell <strong>as</strong>say kits on the market• Reproducibility that comes from using a validated system• Time-saving, no-transfection kit includes cells, media, detectionreagents and controls1. Plate 2. Treat3. Develop4. DetectLeading Light TM CellsGrowth MediumAssay MediumFreezing MediumWnt-3a ProteinDkk-1 ProteinLithium Chloride (Optional)Lucifer<strong>as</strong>e SubstrateLucifer<strong>as</strong>e Assay BufferActivation by Wnt-3a Proteinβ-CateninWnt(Activator)Wnt(Activator)FrizzledLRP5/6Dshβ-Cateninβ-CateninGSK3Frat-1NuclearLocalizationIn-tact Pathway For Screening Flexibility- Small Molecules- Biotherapeutics- Genetically Modified Cells (e.g. transfect, siRNA, etc.)AxinLucifer<strong>as</strong>eLucifer<strong>as</strong>eLucifer<strong>as</strong>ep300/CBPβ-CateninLucifer<strong>as</strong>e ReporterTCF/LEFLeading Light TMInhibition by Dkk-1TCF TCF TCF TCF TCF TCF TATA FIREFLY LUCIFERASEWnt ReporterCell LineWnt(Activator)+Dkk-1(Inhibitor)WntWntWIF-1sFRPFrizzledSOSTProte<strong>as</strong>omaldegradationLRP5/6DkkLRP5/6KremenAPCPβ-CateninGSK3AxinWnt Pathway Compound Library (BML-2838)The Screen-Well ® Wnt Pathway Library offers a convenient solution for benchmarking compounds against a collection of 75known Wnt modulators. The library is available in 100uL and 500uL formats (dissolved in DMSO) for use <strong>as</strong> a companion to theLeading Light Wnt Reporter Assay, or individually <strong>as</strong> a general screening tool for chemical genomics or Wnt <strong>as</strong>say.ScreenWell TM Wnt Pathway Library Targets IncludeWnt5a Dkk Axin Porcupine β-catenin LRP5/6TCF Dishevelled GSK-3B Adenylate Cycl<strong>as</strong>e COX-II sFRP-1
QUANTIFY CRITICAL WNT PATHWAY PROTEINSSensitive Wnt Pathway ELISA KitsObtain fully quantitative data with immunometric detection of β-Catenin, total and phosphorylated GSK-3β, and DKK-1 inhuman, mouse and rat samples.• Sensitive <strong>as</strong>says me<strong>as</strong>ure picogram levels compared to microgram levels in Western blot• Suitable for cell lysates, culture supernates, pl<strong>as</strong>ma, or serum sample types (see table)• High throughput format allows analysis of 40 samples in duplicate in less than 3 hours• Simple protocols and liquid color-coded reagents save time and reduce errorPRODUCT SENSITIVITY SPECIFICITY SAMPLE TYPESDkk-1 Human Mouse Rat Cell Lysates Culture Supernates Serum Pl<strong>as</strong>maADI-900-151 0.98 pg/mL • • • •ADI-900-172 116.7 pg/mL • • • •ADI-900-171 26.1 pg/mL • • • •GSK-3β Human Mouse Rat Cell Lysates Culture Supernates Serum Pl<strong>as</strong>maADI-900-144 74.4 pg/mL • • •[pSer9] GSK-3β Human Mouse Rat Cell Lysates Culture Supernates Serum Pl<strong>as</strong>maADI-900-123A 9.0 pg/mL • • • •β-catenin Human Mouse Rat Cell Lysates Culture Supernates Serum Pl<strong>as</strong>maADI-900-135 27.0 pg/mL • • • •Highly Characterized Wnt Related Proteins & Antibodies<strong>Enzo</strong> offers a wide selection of high-quality reagents for analysis of Wnt related signal transduction, including purifiedproteins and thoroughly validated monoclonal and polyclonal antibodies.Select Wnt Pathway Proteins & AntibodiesProduct Name Product # Product Name Product #C<strong>as</strong>ein Kin<strong>as</strong>e II (human), (recombinant) BML-SE124 Adenomatous polyposis coli (APC), pAb ADI-905-322Dkk-1 (mouse), (recombinant) (His-tag) ADI-KPR-CP100 β-Catenin, mAb (12F7) ADI-KAM-ST001GSK-3β (human), (recombinant) (GST-tag) BML-SE355 Dishevelled-1 pAb BML-DA4170Wnt-3a (mouse), (recombinant) ENZ-60001 Dishevelled-2 pAb BML-DA4270GSK-3α/β, mAb (1H8)ADI-KAM-ST002GSK-3β, pAbADI-KAP-ST002Monitor Wnt signal transductionfrom cell surface to nucleus.www.enzolifesciences.com
Biochem®<strong>Life</strong> <strong>Sciences</strong>scientists enabling scientists.GLOBAL HEADQUARTERS<strong>Enzo</strong> <strong>Life</strong> <strong>Sciences</strong> Inc.10 Executive BoulevardFarmingdale, NY 11735Phone: 1.800.942.0430Fax: 631.694.7501info-usa@enzolifesciences.comTherapeuticsEUROPE/ASIA<strong>Enzo</strong> <strong>Life</strong> <strong>Sciences</strong> (ELS) AGIndustriestr<strong>as</strong>se 17®CH-4415 Lausen, SwitzerlandPhone: +41 61 926 8989Fax: +41 61 926 8979info-eu@enzolifesciences.comFor local distributors and detailed product information visit us online:www.enzolifesciences.comMB7-100112Clinical LabsPut our experience to work for you!Contact <strong>Enzo</strong> to learn more about how our scientists are working to provide innovative tools for your research.
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