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03-119 RIPAb+™ CUGBP2 - RIP Validated Antibody and Primer Set

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03-119
10 assays  10 assays per set. Recommended use: ~5 μg of antibody per RIP (dependent upon biological context).
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey Applications
      HRIP, WB, IP
      Description
      Catalogue Number03-119
      Brand Family Upstate
      Trade Name
      • RIPAb+
      • Upstate
      DescriptionRIPAb+™ CUGBP2 - RIP Validated Antibody and Primer Set
      OverviewRIPAb+ antibodies are evaluated using the RNA Binding Protein Immunoprecipitation (RIP) assay. Each RIPAb+ antibody set includes a negative control antibody to ensure specificity of the RIP reaction and is verified for the co-immunoprecipitation of RNA associated specifically with the immunoprecipitated RNA binding protein of interest. Where appropriate, the RIPAb+ set also includes quantitative RT-PCR control primers (RIP Primers) to biologically validate your IP results by successfully co-precipitating the specific RNA targets, such as messenger RNAs. The qPCR protocol and primer sequences are provided, allowing researchers to validate RIP protocols when using the antibody in their experimental context. If a target specific assay is not provided, the RIPAb+ kit is validated using an automated microfluidics-based assay by enrichment of detectable RNA over control immunoprecipitation.
      CUGBP2 is a RNA-binding protein implicated in the regulation of several post-transcriptional events including pre-mRNA alternative splicing, mRNA translation and stability. CUGBP2 mediates exon inclusion and/or exclusion in pre-mRNA that are subject to tissue-specific and developmentally regulated alternative splicing.
      Background InformationThe survival of motor neurons (SMN) complex is essential for the biogenesis of small nuclear ribonucleoprotein (snRNP) complexes in eukaryotic cells. Reduced levels of SMN cause the motor neuron degenerative disease, spinal muscular atrophy. Complexes containing Gemin3-Gemin4-Gemin5 and Gemin6-Gemin7-unrip persist at similar levels when SMN is reduced. Gemin5-containing subunits bind small nuclear RNA independently of the SMN complex and without a requirement for exogenous ATP. ATP hydrolysis is, however, required for displacement of small nuclear RNAs from the Gemin5-containing subunits and their assembly into snRNPs.
      References
      Product Information
      FormatPurified
      Control
      • Includes negative control mouse IgG antibody and control primers specific for the cDNA of human JUN.
      PresentationAnti-CUGBP2 (Mouse Monoclonal), Part # CS204360. One vial containing 50 µg of protein A purified mouse monoclonal IgG1 in 50 µL of 1X PBS, pH 7.0, and 0.1% sodium azide, before the addition of 30% glycerol. Store at -20°C.
      Normal Mouse IgG, Part # CS200621. One vial containing 125 µg of purified mouse IgG in 125 µL of storage buffer containing 0.1% sodium azide. Store at -20°C.
      RIP Primers, JUN, Part # CS203198.
      One vial containing 75 μL of 5 μM of each primer specific for the cDNA of human JUN.
      Store at -20°C.
      FOR: TCG ACA TGG AGT CCC AGG A
      REV: GGC GAT TCT CTC CAG CTT CC
      Quality LevelMQ100
      Applications
      ApplicationThis RIPAb+ CUGBP2 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • RNA Binding Protein Immunoprecipitation (RIP)
      • Western Blotting
      • Immunoprecipitation
      Application NotesImmunoprecipitation from RIP lysate:
      Representative lot data.
      RIP lysate from Jurkat cells (~5 X 10E7 cell equivalents per IP) was subjected to immunoprecipitation using 5 µg of either a normal mouse IgG, (Cat. #CS200621), or 5 µg of Anti-CUGBP2 antibody (Cat. # CS204360).
      Precipitated proteins (lane 1: mouse IgG, lane 2: anti-CUGBP2) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-CUGBP2 antibody (Cat. # CS204360, 1:1000). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00231)
      Arrow indicates CUGBP2 (Figure 2).
      Western Blot Analysis:
      Representative lot data.
      Lysates from HL60 cells were probed with a previous lot of Anti-CUGBP2 (1:1000 dilution).
      Arrow indicates CUGBP2 ~56 kDa (Figure 3).
      Biological Information
      ImmunogenA synthetic peptide corresponding to amino acids 209-415 of human CuGBP2.
      ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
      HostMouse
      SpecificityCuGBP2
      IsotypeIgG1
      Species Reactivity
      • Human
      Species Reactivity NoteHuman. Other species have not yet been tested.
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryMembers of the CELF/BRUNOL protein family contain two N-terminal RNA recognition motif (RRM) domains, one C-terminal RRM domain, and a divergent segment of 160-230 aa between the second and third RRM domains. Members of this protein family regulate pre-mRNA alternative splicing and may also be involved in mRNA editing, and translation. Alternative splicing results in multiple transcript variants encoding different isoforms.
      Gene Symbol
      • CUGBP2
      • CUG-BP2
      • BRUNOL3
      • CELF2
      • CELF-2
      • Etr-3
      • ETR-3
      • ETR3
      • NAPOR-2
      • NAPOR2
      • NAPOR
      • hNAPOR
      Purification MethodProtein A Purfied
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: O95319 # RNA-binding protein implicated in the regulation of several post-transcriptional events. Involved in pre-mRNA alternative splicing, mRNA translation and stability. Mediates exon inclusion and/or exclusion in pre-mRNA that are subject to tissue-specific and developmentally regulated alternative splicing. Specifically activates exon 5 inclusion of TNNT2 in embryonic, but not adult, skeletal muscle. Activates TNNT2 exon 5 inclusion by antagonizing the repressive effect of PTB. Acts as both an activator and repressor of a pair of coregulated exons: promotes inclusion of the smooth muscle (SM) exon but exclusion of the non-muscle (NM) exon in actinin pre-mRNAs. Promotes inclusion of exonS 21 and exclusion of exon 5 of the NMDA receptor R1 pre- mRNA. Involved in the apoB RNA editing activity. Increases COX2 mRNA stability and inhibits COX2 mRNA translation in epithelial cells after radiation injury (By similarity). Modulates the cellular apoptosis program by regulating COX2-mediated prostaglandin E2 (PGE2) expression (By similarity). Binds to (CUG)n triplet repeats in the 3'-UTR of transcripts such as DMPK. Binds to the muscle-specific splicing enhancer (MSE) intronic sites flanking the TNNT2 alternative exon 5. Binds preferentially to UG-rich sequences, in particular UG repeat and UGUU motifs. Binds to apoB mRNA, specifically to AU-rich sequences located immediatly upstream of the edited cytidine. Binds AU-rich sequences in the 3'-UTR of COX2 mRNA (By similarity). Binds to an intronic RNA element responsible for the silencing of exon 21 splicing (By similarity). Binds to (CUG)n repeats (By similarity).


      SIZE: 508 amino acids; 54285 Da

      SUBUNIT: Found in a complex with APOBEC1 and ACF.

      SUBCELLULAR LOCATION: Nucleus. Cytoplasm. Note=Accumulates in the cytoplasm after ionizing radiation (By similarity). Colocalizes with APOBEC1 and ACF. RNA-binding activity is detected in both nuclear and cytoplasmic compartments.

      TISSUE SPECIFICITY: Expressed in frontal cortex. Isoform 1 is expressed in brain and lung. Isoform 2 is expressed in heart, brain, placenta, lung, liver, kidney, skeletal muscle and pancreas. Isoform 4 is expressed in heart, lung, skeletal muscle, kidney and pancreas.
      DEVELOPMENTAL STAGE: Isoform 1 is expressed in fetal brain. Isoform 2 is expressed in fetal heart, brain, thymus, lung, liver, skeletal muscle, kidney and spleen. Isoform 4 is expressed in fetal heart, brain, thymus, lung and skeletal muscle.




      SIMILARITY: SwissProt: O95319 ## Belongs to the CELF/BRUNOL family. & Contains 3 RRM (RNA recognition motif) domains.
      Molecular Weight56 kDa
      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 Jurkat cells (~5 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal mouse IgG (Cat. # CS200621), or 5 µg of Anti-CUGBP2 antibody (Cat. # CS204360) and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
      Successful immunoprecipitation of CUGBP2-associated RNA was verified by qPCR using RIP Primers Jun, (Cat. # CS203198) (Figure 1).
      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 receipt, 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 Size10 assays
      Material Package10 assays per set. Recommended use: ~5 μg of antibody per RIP (dependent upon biological context).
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      RIPAb+™ CUGBP2 - RIP Validated Antibody and Primer Set SDS

      Title

      Safety Data Sheet (SDS) 

      RIPAb+™ CUGBP2 - RIP Validated Antibody and Primer Set Certificates of Analysis

      TitleLot Number
      RIPAb+ CUGBP2 - NRG1804157 NRG1804157
      RIPAb+™ CUGBP2 - 3563625 3563625
      RIPAb+™ CUGBP2 -2544325 2544325
      RIPAb+™ CUGBP2 -2736384 2736384
      RIPAb+™ CUGBP2 -2872386 2872386

      References

      Reference overviewPub Med ID
      Expression of transcription factors and matrix genes in response to serum stimulus in vascular smooth muscle cells.
      André Markmann,Jürgen Rauterberg,Peter Vischer,Horst Robenek,Frank Echtermeyer,Heike Will,Daniela G Seidler,Marian F Young,Hans Kresse
      European journal of cell biology  82  2003

      Show Abstract
      12691261 12691261

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      Categories

      Life Science Research > Antibodies and Assays > Immunoassays > Immunoprecipitation (IP) > RNA-Binding Protein Immunoprecipitation (RIP) > RIP Validated Antibodies
      Life Science Research > Antibodies and Assays > Primary Antibodies
      Life Science Research > Protein Detection and Quantification > Immunoassays > Immunoprecipitation (IP) > RNA-Binding Protein Immunoprecipitation (RIP) > RIP Validated Antibodies