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03-901 RIPAb+™ FLAG - RIP Validated Antibody and Primer Set

This RIPAb+™ FLAG -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Research Category: Epigenetics & Nuclear Function
03-901
1 Kit  
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      Overview

      Replacement Information

      Key Spec Table

      Species Reactivity
      E. coli, Vrt
      Description
      Catalogue Number03-901
      DescriptionRIPAb+™ FLAG - RIP Validated Antibody and Primer Set
      Background InformationEpitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance or poorly immunogenic proteins when protein specific antibodies are not available. Tagging with FLAG may be done at the N-terminus, N-terminus preceded by a methionine residue, C-terminus, and in internal positions of the target protein. The small size of the epitope tag and its high hydrophilicity tend to decrease the possibility of interference with protein expression, proteolytic maturation, antigenicity and function. The enterokinase cleavage site allows it to be completely removed from the purified fusion proteins.
      References
      Product Information
      Components
      • Anti-FLAG M2 (mouse monoclonal IgG1): Part No. CS226432
        Normal Mouse IgG: Part No. CS200621
        RIP Primers, U1 snRNA: Part # CS203215
      FormatPurified
      PresentationAnti-FLAG M2: (mouse monoclonal IgG1): One vial containing 50 μg of protein G purified antibody in 50 μL buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4). Store at -20°C.
      Normal Mouse IgG: One vial containing 125 μg purified mouse IgG in 125 μL storage buffer containing 0.1% sodium azide. Store at -20°C.
      RIP Primers, U1 snRNA: One vial containing 75 μL of 5 μM of each primer specific for the cDNA of human U1 snRNA. Store at -20°C.
      FOR: GGG AGA TAC CAT GAT CAC GAA GGT
      REV: CCA CAA ATT ATG CAG TCG AGT TTC CC
      Quality LevelMQ200
      Applications
      ApplicationThis RIPAb+™ FLAG -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Application NotesRNA Binding Protein Immunoprecipitation:
      RIP Lysate prepared from HeLa cells (1 X 10E7 cell equivalents per IP) expressing SNRNP70-GFP-FLAG (Cat. No. 03-903) by RNA transfection was subjected to immunoprecipitation using 5 µg of either a normal mouse IgG or Anti-FLAG M2 antibody and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700). Successful immunoprecipitation of SNRNP70-GFP-FLAG associated RNA was verified by qPCR using RIP Primers, U1 snRNA.

      Immunoprecipitation:
      RIP Lysate prepared from HeLa cells (1 X 10E6 cell equivalents per IP) expressing SNRNP70-GFP-FLAG (Cat. No. 03-903) by RNA transfection was subjected to immunoprecipitation using 0.5 µg of either a normal rabbit IgG, or Anti-FLAG M2 antibody. Precipitated proteins were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-FLAG M2 (1.0 µg/mL). Proteins were visualized using Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP (Cat. # AP124P) and a chemiluminescence detection system.
      Biological Information
      ImmunogenFLAG; peptide sequence DYKDDDDK
      CloneM2
      ConcentrationPlease refer to product datasheet.
      HostMouse
      SpecificityDYKDDDDK sequence (FLAG) containing proteins. Reacts with both N-terminal and C-terminal fusion proteins.
      IsotypeIgG
      Species Reactivity
      • E. coli
      • Vertebrates
      Antibody TypeMonoclonal Antibody
      Purification MethodProtein G purified
      Molecular WeightVarious
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceRNA Binding Protein Immunoprecipitation: RIP Lysate prepared from HeLa cells (1 X 10E7 cell equivalents per IP) expressing SNRNP70-GFP-FLAG (Cat. No. 03-903) by RNA transfection was subjected to immunoprecipitation using 5 µg of either a normal mouse IgG or Anti-FLAG M2 antibody and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700). Successful immunoprecipitation of SNRNP70-GFP-FLAG associated RNA was verified by qPCR using RIP Primers, U1 snRNA.
      Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.
      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 ConditionsStable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
      Packaging Information
      Material Size1 Kit
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      03-901 04065265138680