03-102 RIPAb+ HuR - RIP Validated Antibody and Primer Set

03-102
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
      H, M, R RIP, IP, WB, ICC, IHC
      Description
      Catalogue Number03-102
      Brand Family Upstate
      Trade Name
      • RIPAb+
      • Upstate
      DescriptionRIPAb+ HuR - RIP Validated Antibody and Primer Set
      OverviewRIPAb+ antibodies are evaluated using the RNA Binding Protein Immunoprecipitation (RIP) assay. Each RIPAb+ antibody set includes quantitative RT-PCR control primers (RIP Primers) to biologically validate your IP results by successfully coprecipitating a specific RNA target (where such a specific target is known). The qPCR protocol and primer sequences are provided, allowing researchers to validate RIP protocols when using the antibody in their experimental context. Each set also includes a negative control antibody to ensure specificity of the RIP reaction.
      The RIPAb+ HuR set includes the HuR antibody, a negative control antibody (purified Rabbit IgG), and positive qPCR primers which amplify a 100 bp fragment of the human beta actin (ACTB) cDNA. The HuR and negative control antibodies are supplied in a scalable "per RIP" reaction size and can be used to functionally validate the precipitation HuR-associated RNAs.
      Alternate Names
      • ELAV-like protein 1
      • Hu-antigen R
      Background InformationHu-antigen R (HuB), ELAV-like protein 1, (ELAV1) is a transcription factor belonging to the ELAV family of RNP proteins. It binds avidly to the Au-rich element in cFos and interleukin-3 mRNAs.
      References
      Product Information
      FormatPurified
      Control
      • Included negative control rabbit IgG antibody and control primers specific for human beta actin (ATCB).
      PresentationAnti-HuR (rabbit polyclonal). One vial containing 50 μg of purified rabbit polyclonal in 100 μL of buffer containing PBS in 0.05% sodium azide with 50% Glycerol. Liquid at -20°C.

      Normal Rabbit IgG. One vial containing 125 μg purified rabbit IgG in 125 μL storage buffer containing 0.1% sodium azide. Store at -20°C.

      RIP Primers, ACTB. One vial containing 75 μL of 5 μM of each primer specific for human beta actin (ACTB). Store at -20°C.
      FOR: TTG TTA CAG GAA GTC CCT TGC C
      REV: ATG CTA TCA CCT CCC CTG TGT G
      Applications
      ApplicationThis RIPAb+ HuR -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • RNA Binding Protein Immunoprecipitation (RIP)
      • Immunoprecipitation
      • Western Blotting
      • Immunocytochemistry
      • Immunohistochemistry
      Application NotesImmunoprecipitation from RIP lysate:
      RIP lysate from MCF7 cells (2 X 106 cell equivalents per IP) was subjected to immunoprecipitation using 0.5 μg of either a normal rabbit IgG or Anti-HuR antibody. Precipitated proteins were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-HuR (1.0 μg/mL).
      Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00231) (Please see figures).

      Western Blot Analysis:
      3T3/A31 lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with HuR, hu (1:500 dilution). Proteins were visualized using a Donkey anti-Rabbit conjugated to HRP and visualized using a chemiluminescence detection system. (Please see figures).

      Immunoprecipitation Analysis:
      10 ug of this antibody immunoprecipitated HuR from 500 ug of 3T3/A31 RIPA lysate (Please see figures).

      mmunocytochemistry Analysis:
      Confocal IF analysis of HeLa, NIH/3T3 using anti-HuR (Red). Actin filaments have been labeled with AlexaFluor® 488 -Phalloidin (Green). Nucleus is stained with DAPI (Blue) (Please see figures).
      Biological Information
      ImmunogenKLH-conjugated, synthetic peptide corresponding to amino acids 1-13 of human HuR.
      Epitopea.a. 1-13 of HuR
      HostRabbit
      SpecificityHuR
      Species Reactivity
      • Human
      • Mouse
      • Rat
      Antibody TypePolyclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryThe protein encoded by this gene is a member of the ELAVL protein family. This encoded protein contains 3 RNA-binding domains and binds cis-acting AU-rich elements. It destabilizes mRNAs and thereby regulates gene expression.
      Gene Symbol
      • ELAVL1
      • MelG
      • HUR
      • ELAV1
      • HuR
      • Hua
      Purification MethodProtein A Purfied
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: Q15717 # Binds avidly to the AU-rich element in FOS and IL3/interleukin-3 mRNAs. In the case of the FOS AU-rich element, HUR binds to a core element of 27 nucleotides that contain AUUUA, AUUUUA, and AUUUUUA motifs.
      SIZE: 326 amino acids; 36092 Da
      SUBUNIT: Interacts with ANP32A.
      TISSUE SPECIFICITY: Ubiquitous.
      PTM: Methylated at Arg-217 by CARM1 in macrophages in response to LPS challenge.
      SIMILARITY: SwissProt: Q15717 ## Belongs to the RRM elav family. & Contains 3 RRM (RNA recognition motif) domains.
      Molecular Weight35 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 HeLa cells (2 x 107 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal Rabbit IgG or Anti-HuR antibody and the Magna RIP Kit (Cat. # 17-700).
      Successful immunoprecipitation of HuR-associated RNA was verified by qPCR using RIP Primers, ACTB (Please see figures).
      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.
      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

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      RIPAb+ HuR3031910
      RIPAb+ HuR - 21052212105221
      RIPAb+ HuR - 24333482433348
      RIPAb+ HuR - 24590522459052
      RIPAb+ HuR - 20008292000829
      RIPAb+ HuR - 21996422199642
      RIPAb+ HuR - 24889012488901
      RIPAb+ HuR - 31868123186812
      RIPAb+ HuR - JH1775074JH1775074
      RIPAb+ HuR - NG1919889NG1919889

      References

      Reference overviewApplicationPub Med ID
      RNA-binding proteins regulate the expression of the immune activating ligand MICB.
      Nachmani, D; Gutschner, T; Reches, A; Diederichs, S; Mandelboim, O
      Nature communications  5  4186  2014

      Show Abstract
      Western Blotting24924487 24924487
      Heat resistance of Bacillus spores when adhered to stainless steel and its relationship to spore hydrophobicity.
      P Simmonds,B L Mossel,T Intaraphan,H C Deeth
      Journal of food protection  66  2003

      Show Abstract
      14627285 14627285

      Brochure

      Title
      RNA-Binding Protein Immunoprecipitation

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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