03-107 | RIPAb+ EF1α - RIP Validated Antibody and Primer Set

03-107
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 Spec Table

      Species ReactivityKey Applications
      H, Ma, Yeast (S. cerevisiae)RIP, IP, WB
      Description
      Catalogue Number03-107
      Brand Family Upstate
      Trade Name
      • RIPAb+
      • Upstate
      DescriptionRIPAb+ EF1α - 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.
      Alternate Names
      • Elongation factor 1-alpha 1
      • Eukaryotic elongation factor 1 A-1
      • Elongation factor Tu
      • Leukocyte receptor cluster member 7
      Background InformationHuman eukaryotic translation elongation factor 1 alpha has two isoforms, alpha 1 and alpha 2. The alpha 1 isoform gene encodes a subunit of the elongation factor-1 complex which is responsible for the enzymatic delivery of aminoacyl tRNAs to the ribosome. The alpha 1 isoform is expressed in brain, placenta, lung, liver, kidney, and pancreas. The other isoform (alpha 2) is expressed in brain, heart and skeletal muscle.
      The alpha 1 isoform is identified as an autoantigen in 66% of patients with Felty syndrome. The gene for the alpha 1 isoform has been found to have multiple copies on many chromosomes, some of which, if not all, represent different pseudogenes. The gene for the alpha 2 may be critical in the development of ovarian cancer.
      References
      Product Information
      FormatPurified
      Control
      • Includes negative control mouse IgG antibody.
      PresentationAnti-EF1α (Mouse Monoclonal IgG1κ). One vial containing 50 μg of protein G purified mouse monoclonal IgG1κ in 72 μL of 70% storage buffer containing (0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide) and 30% glycerol. Store at -20°C.

      Normal Mouse IgG.
      One vial containing 125 μg of purified mouse IgG in 125 µL of storage buffer containing 0.1% sodium azide. Store at -20°C.
      Applications
      ApplicationThis RIPAb+ EF1α -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • RNA Binding Protein Immunoprecipitation (RIP)
      • Immunoprecipitation
      • Western Blotting
      Application NotesImmunoprecipitation from RIP Lysate:
      Representative lot data.
      RIP lysate from HeLa cells (2 X 106 cell equivalents per IP) was subjected to immunoprecipitation using 0.5 µg of either a normal mouse IgG or Anti-EF1α antibody. Precipitated proteins (Lane 1: mouse IgG, Lane 2: anti-EF1α) and HeLa whole cell lysate (Lane 3) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-EF1α antibody (1.0 µg/mL).
      Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00232) (please see figures).

      Automated Microfluidics-based Electrophoretic RNA Separation and Analysis:
      RIP Lysate prepared from HeLa cells (2 X 107 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal mouse IgG or Anti-EF1α antibody and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
      Successful immunoprecipitation of EF1a-associated RNA was verified by automated microfluidics-based electrophoretic RNA separation and analysis. Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.

      Electropherograms were generated by plotting fluorescence intensities versus migration times.
      (Please see figures)

      The virtual gel view was created from this data.
      (Please see figures)
      Biological Information
      ImmunogenCrude calmodulin-binding proteins from Trypanosoma brucei isolated using calmodulin-affinity chromatography.
      HostMouse
      SpecificityEF1α (Elongation factor-1 alpha)
      IsotypeIgG1κ
      Species Reactivity
      • Human
      • Mammals
      • Yeast (S. cerevisiae)
      Species Reactivity NoteHuman. Predicted to react with most mammals, and yeast.
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryThis gene encodes an isoform of the alpha subunit of the elongation factor-1 complex, which is responsible for the enzymatic delivery of aminoacyl tRNAs to the ribosome. This isoform (alpha 1) is expressed in brain, placenta, lung, liver, kidney, and pancreas, and the other isoform (alpha 2) is expressed in brain, heart and skeletal muscle. This isoform is identified as an autoantigen in 66% of patients with Felty syndrome. This gene has been found to have multiple copies on many chromosomes, some of which, if not all, represent different pseudogenes.
      Gene Symbol
      • EEF1A1
      • EEF1A
      • EF1A
      • LENG7
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P68104 # This protein promotes the GTP-dependent binding of aminoacyl-tRNA to the A-site of ribosomes during protein biosynthesis.
      SIZE: 462 amino acids; 50141 Da
      SUBUNIT: Found in a nuclear export complex with XPO5, EEF1A1, Ran and aminoacylated tRNA. Interacts with XPO5.
      SUBCELLULAR LOCATION: Cytoplasm.
      TISSUE SPECIFICITY: Brain, placenta, lung, liver, kidney, pancreas but barely detectable in heart and skeletal muscle.
      SIMILARITY: SwissProt: P68104 ## Belongs to the GTP-binding elongation factor family. EF-Tu/EF-1A subfamily.
      Molecular Weight53 kDa
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceFluorescence-based Quantifiation Assay:
      Successful immunoprecipitation of EF1α-associated RNA was verified by enrichment over mock immunoprecipitation using a fluorescence-based quantification assay.
      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. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
      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

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      Categories

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