17-10521 EZ-Magna Nuclear RIP™ (Cross-Linked)
Nuclear RNA-Binding Protein Immunoprecipitation Kit

17-10521
24 assays  Kit capacity: 24 RNA-binding protein immunoprecipitation assays using a cross-linked nuclear lysate
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      Overview

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      Key Spec Table

      Key Applications
      RIP, Protein Interaction Assays
      Description
      Catalogue Number17-10521
      Trade Name
      • Magna Nuclear RIP
      DescriptionEZ-Magna Nuclear RIP™ (Cross-Linked)
      Nuclear RNA-Binding Protein Immunoprecipitation Kit
      OverviewFeatures and Advantages

      • Generates cross-linked chromatin to allow analysis of a variety of chromatin-associated RNAs
      • Flexible, scalable input requirements: Recover RNA from millions of cells or as few as 5,000 cells
      • Magnetic protein A/G bead blend and optimized buffer system results in low backgrounds and high signal-to-noise ratios
      • Suitable for analysis by RT-qPCR or RIP-seq
      • Complete set of reagents and detailed protocol to enable first-time success


      Magna Nuclear RIP™ Kits are specially designed to allow the discovery and analysis of a variety of chromatin associated RNAs such as long non-coding RNAs, enhancer RNAs and miRNAs . These chromatin-associated RNAs often regulate gene expression and can be analyzed with applications including quantitative reverse transcription polymerase chain reaction (RT-PCR), microarray analysis (RIP-chip) and next generation sequencing (RIP-Seq).

      Nuclear RIP can be performed using chromatin that has interactions stabilized by formaldehyde treatment (cross-linked) or chromatin that has not been treated with a cross linking reagent (native). While both of these approaches are similar in that they are designed to recover chromatin associated RNA, the reagents used and the details of the protocol and types of interactions typically detected are different. Cross-linked can capture higher molecular weight complexes in in vivo configurations with possibly lower affinities. In contrast native RIP is expected to recover high affinity, more direct interactions between proteins encoded RNA binding motifs and candidate RNAs. For less well understood proteins and protein complexes often both approaches are used.

      The kit described here is for the cross-linked approach. If a native approach is of interest please visit the product page for the Magna Nuclear RIP (Native) Kit, catalogue # 17-10522 or the EZ-Magna Nuclear RIP (Native) Kit, catalogue # 17-10523.
      Materials Required but Not DeliveredReagents
      • Cells, stimulated or treated as needed for the experimental system
      • 37% formaldehyde
      • Antibody of interest for RNA-binding protein immunoprecipitation (RIP)
      • Negative Control Antibody
      • PBS (RNase free)
      • TRIzol® LS Reagent (Life Technologies Cat. #10296-010)
      • Chloroform (e.g. Fisher, Cat. # BP1145)
      • 100% Ethanol (molecular biology grade)
      • Isopropanol (molecular biology grade)
      • Precipitation Carrier (e.g. Pellet Paint® Co-Precipitant (125 reactions, EMD Millipore Cat. # 69049) or linear Acrylamide (5mg/mL) if samples are intended for Next Gen Sequence library preparation.
      • Nuclease Free Water
      • RNase A (10 mg/mL)
      • Liquid nitrogen (Optional)
      • Reagents for qRT-PCR Analysis and/or Reagents for RNA-Seq Library Construction (optional)


      Equipment
      • Dounce homogenizer (loose pestle, necessary for tissue samples but optional for cultured cells)
      • Sonicator
      • Magnetic Separator
      • Magna GrIP™ Rack (8 well, Millipore Cat. # 20-400) or PureProteome™ Magnetic Stand, (Millipore Cat. # LSKMAGS08)
      • Vortex mixer
      • Rotating wheel/platform
      • Microcentrifuge
      • -80°C freezer
      • Thermomixer (60°C capable)
      • Variable temperature water bath or incubator
      • Rotating microtube mixer
      • Real-time PCR thermal cycler
      References
      Product Information
      Components
      • 10X Glycine
      • 10X PBS
      • Nuclei Isolation Buffer
      • RIP Cross-Linked Lysis Buffer
      • Protein A/G Magnetic Beads
      • Nuclear RIP Dilution Buffer
      • Low Salt Wash Buffer
      • High Salt Wash Buffer
      • LiCl Wash Buffer
      • TE Buffer
      • RIP Elution Buffer
      • 10% SDS
      • 0.5 M EDTA
      • DNase I (RNase Free)
      • DNase I Supplement
      • DNase I Reaction Buffer
      • Protease Inhibitor Cocktail III, Animal Free
      • RNAse Inhibitor
      • Proteinase K

      • Control Antibodies and Primers
        Normal Mouse IgG Negative Control Antibody
      • Anti-EZH2 Positive Control Antibody
      • NEAT1 Positive Control Primers
      • U1 snRNA Negative Control Primers
      PresentationTwo boxes containing key reagents for generation of cross linked nuclear lysates and performance of 24 individual RNA-binding protein immunoprecipitation (RIP) reactions. Plus box containing positive and negative control antibodies and qPCR primers.
      Applications
      ApplicationEZ-Magna Nuclear RIP (Cross-Linked) RNA-Binding Protein Immunoprecipitation Kit is designed for the analysis of chromatin associated RNA such lncRNAs, enhancer RNAs and miRNAs.
      Key Applications
      • RNA Binding Protein Immunoprecipitation (RIP)
      • Protein Interaction Assays
      Biological Information
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      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
      Storage ConditionsUpon receipt, store components at the temperatures indicated on the labels.
      Kit components are stable for 6 months from date of shipment when stored as directed.
      Packaging Information
      Material Size24 assays
      Material PackageKit capacity: 24 RNA-binding protein immunoprecipitation assays using a cross-linked nuclear lysate
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      EZ-Magna Nuclear RIP™ (Cross-Linked)
      Nuclear RNA-Binding Protein Immunoprecipitation Kit SDS

      Title

      Safety Data Sheet (SDS) 

      Technical Info

      Title
      Effective Nuclear RIP Protocols to Study Interactions between Proteins and Non-Coding RNAs
      White Paper - The Message in the Marks: Deciphering Cancer Epigenetics

      User Guides

      Title
      Magna Nuclear RIP¿ (Cross-Linked) Nuclear RNA-Binding Protein Immunoprecipitation Kit

      Instructions for Use

      Title
      EZ-Nuclear RIP (Cross-Linked) Nuclear RNA-Binding Protein Immunoprecipitation Kit