03-110 | RIPAb+ Ago2 - RIP Validated Antibody and Primer Set

03-110
10 assays  10 assays per set. Recommended use: ~5 μL of antibody per RIP (dependent upon biological context).
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      Overview

      Replacement Information

      Key Spec Table

      Species ReactivityKey Applications
      H, BRIP, IP, WB
      Description
      Catalogue Number03-110
      Brand Family Upstate
      Trade Name
      • RIPAb+
      • Upstate
      DescriptionRIPAb+ Ago2 - 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
      • Argonaute-2
      • Eukaryotic translation initiation factor 2C
      • Slicer protein
      • Piwi domain protein
      Background InformationAgo2 (argonaute-2), also known as eukaryotic translation initiation factor 2C (EIF2C2), is an essential component for siRNA-directed RNA interference (RNAi) response. Ago2 is an endonuclease required for the unwinding of siRNA duplex and assembly of siRNA into RNA-induced silencing complexes (RISC). Ago2 interacts with DICER1 through its Piwi domain. This Piwi domain is thought to provide RNA cleavage activity via a mechanism similar to RNase H. Ago2 activity is necessary for embryonic development as well as RNA-mediated gene silencing (RNAi).
      References
      Product Information
      FormatAscites
      Control
      • Includes negative control mouse IgG antibody and control primers specific for human FOS.
      PresentationAnti-Ago2 (Mouse Monoclonal IgG1κ). One vial containing 50 µL of ascites with 0.05% sodium azide. Store at -20°C.

      Normal Mouse IgG. One vial containing 125 μg of purified mouse IgG in 125 µL of stoage buffer containing 0.1% sodium azide. Store at -20°C.

      RIP Primers, FOS. One vial containing 75 μL of 5 μM of each primer specific for human FOS gene. Store at -20°C.
      FOR: GAG AGC TGG TAG TTA GTA GCA TGT TGA
      REV: AAT TCC AAT AAT GAA CCC AAT AGA TTA GTT A
      Applications
      ApplicationThis RIPAb+ Ago2 -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 NotesRNA Binding Protein Immunoprecipitation (microRNA):
      RIP Lysate prepared from HeLa cells (2 X 107 cell equivalents per IP) were subjected to immunoprecipitation using 5 µL of either a normal mouse IgG or 5 µL of Anti-Ago2 antibody and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
      Successful immunoprecipitation of Ago2-associated microRNA was verified by TaqMan® microRNA assay, hsa-miR-23a. (Please see figures).
      Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.


      Immunoprecipitation 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 µL of either a normal mouse IgG or Anti-Ago2 antibody. Precipitated proteins (lane 1: mouse IgG, lane 2: anti-Ago2) and HeLa whole cell lysate (lane 3) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Ago2 antibody (1:5,000 dilution).
      Proteins were visualized using goat anti-mouse secondary antibody conjugated to HRP and chemiluminescence detection.
      (Please see figures).

      Biological Information
      ImmunogenKLH-conjugated, synthetic peptide containing the sequence YSGAGPALAPPAPPPPIQG.
      HostMouse
      SpecificityRecognizes Ago2.
      IsotypeIgG1κ
      Species Reactivity
      • Human
      • Bovine
      Species Reactivity NoteHuman. Predicted to react with bovine Ago2 based on sequence homology.
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryThis gene encodes a member of the Argonaute family of proteins which play a role in RNA interference. The encoded protein is highly basic, and contains a PAZ domain and a PIWI domain. It may interact with dicer1 and play a role in short-interfering-RNA-mediated gene silencing. Multiple transcript variants encoding different isoforms have been found for this gene.
      Gene Symbol
      • Ago2
      • eIF-2C 2
      • eIF2C 2
      • PPD
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P45973 # Component of heterochromatin. Recognizes and binds histone H3 tails methylated at 'Lys-9', leading to epigenetic repression. Can interact with lamin B receptor (LBR). This interaction can contribute to the association of the heterochromatin with the inner nuclear membrane. Involved in the formation of functional kinetochore through interaction with MIS12 complex proteins.
      SIZE: 191 amino acids; 22225 Da
      SUBUNIT: Interacts with SUV420H1 and SUV420H2 (By similarity). Interacts directly with ATRX, CHAF1A, LBR, NIPBL, SP100, STAM2 and TRIM28 via the chromoshadow domain. Can interact directly with CBX3 via the chromoshadow domain. Interacts with histone H3 methylated at 'Lys-9'. Interacts with MIS12 and C20orf127.
      SUBCELLULAR LOCATION: Nucleus. Centromere. Note=Component of centromeric and pericentromeric heterochromatin. Associates with chromosomes during mitosis. Associates specifically with chromatin during metaphase and anaphase.
      PTM: Phosphorylation of HP1 and LBR may be responsible for some of the alterations in chromatin organization and nuclear structure which occur at various times during the cell cycle (By similarity). Phosphorylated during interphase and possibly hyper- phosphorylated during mitosis.
      SIMILARITY: SwissProt: P45973 ## Contains 2 chromo domains.
      Molecular Weight100 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 µL of either a normal mouse IgG or 5 µL of Anti-Ago2 antibody and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
      Successful immunoprecipitation of Ago2-associated RNA was verified by qPCR using RIP Primers FOS (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.
      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 μL 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+ Ago2 - 2327500 2327500
      RIPAb+ Ago2 - 2382636 2382636
      RIPAb+ Ago2 - 2444084 2444084
      RIPAb+ Ago2 - 1987489 1987489
      RIPAb+ Ago2 - 2020308 2020308
      RIPAb+ Ago2 - 2064825 2064825
      RIPAb+ Ago2 - 2121817 2121817
      RIPAb+ Ago2 - 2199651 2199651
      RIPAb+ Ago2 - 2287223 2287223
      RIPAb+ Ago2 - 2488822 2488822

      References

      Reference overviewPub Med ID
      Argonaute 2 sustains the gene expression program driving human monocytic differentiation of acute myeloid leukemia cells.
      Iosue, I; Quaranta, R; Masciarelli, S; Fontemaggi, G; Batassa, EM; Bertolami, C; Ottone, T; Divona, M; Salvatori, B; Padula, F; Fatica, A; Lo-Coco, F; Nervi, C; Fazi, F
      Cell death & disease  4  e926  2013

      Show Abstract
      24263100 24263100
      Characterization of polysaccharide production of haemophilus influenzae Type b and its relationship to bacterial cell growth.
      Mickie Takagi,Joaquin Cabrera-Crespo,Júlia Baruque-Ramos,Teresa Cristina Zangirolami,Isaias Raw,Martha Massako Tanizaki
      Applied biochemistry and biotechnology  110  2003

      Show Abstract
      14515024 14515024

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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