Recombinant
RabMAb

Anti-Argonaute-2 antibody [EPR10410] (ab156870)

Overview

  • Product name
    Anti-Argonaute-2 antibody [EPR10410]
    See all Argonaute-2 primary antibodies
  • Description
    Rabbit monoclonal [EPR10410] to Argonaute-2
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Argonaute-2 aa 1-100. The exact sequence is proprietary.

  • Positive control
    • WB: HeLa, MCF7, HepG2 and K562 cell lysates. Rat liver and mouse liver lysates. IHC-P: Human breast carcinoma and human kidney tissues. Flow Cyt: HeLa cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab156870 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/10000. Predicted molecular weight: 97 kDa.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

The mouse and rat recommendation is based on the WB results. This antibody may not be suitable for IHC with mouse or rat samples.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

ICC/IF 1/100 - 1/250.
Flow Cyt 1/200.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function
      Required for RNA-mediated gene silencing (RNAi) by the RNA-induced silencing complex (RISC). The 'minimal RISC' appears to include EIF2C2/AGO2 bound to a short guide RNA such as a microRNA (miRNA) or short interfering RNA (siRNA). These guide RNAs direct RISC to complementary mRNAs that are targets for RISC-mediated gene silencing. The precise mechanism of gene silencing depends on the degree of complementarity between the miRNA or siRNA and its target. Binding of RISC to a perfectly complementary mRNA generally results in silencing due to endonucleolytic cleavage of the mRNA specifically by EIF2C2/AGO2. Binding of RISC to a partially complementary mRNA results in silencing through inhibition of translation, and this is independent of endonuclease activity. May inhibit translation initiation by binding to the 7-methylguanosine cap, thereby preventing the recruitment of the translation initiation factor eIF4-E. May also inhibit translation initiation via interaction with EIF6, which itself binds to the 60S ribosomal subunit and prevents its association with the 40S ribosomal subunit. The inhibition of translational initiation leads to the accumulation of the affected mRNA in cytoplasmic processing bodies (P-bodies), where mRNA degradation may subsequently occur. In some cases RISC-mediated translational repression is also observed for miRNAs that perfectly match the 3' untranslated region (3'-UTR). Can also upregulate the translation of specific mRNAs under certain growth conditions. Binds to the AU element of the 3'-UTR of the TNF (TNF-alpha) mRNA and upregulates translation under conditions of serum starvation. Also required for transcriptional gene silencing (TGS), in which short RNAs known as antigene RNAs or agRNAs direct the transcriptional repression of complementary promoter regions.
    • Sequence similarities
      Belongs to the argonaute family. Ago subfamily.
      Contains 1 PAZ domain.
      Contains 1 Piwi domain.
    • Domain
      The Piwi domain may perform RNA cleavage by a mechanism similar to that of RNase H. However while RNase H utilizes a triad of Asp-Asp-Glu (DDE) for metal ion coordination, this protein appears to utilize a triad of Asp-Asp-His (DDH).
    • Post-translational
      modifications
      Hydroxylated. 4-hydroxylation appears to enhance protein stability but is not required for miRNA-binding or endonuclease activity.
    • Cellular localization
      Cytoplasm > P-body. Nucleus. Translational repression of mRNAs results in their recruitment to P-bodies. Translocation to the nucleus requires IMP8.
    • Information by UniProt
    • Database links
    • Alternative names
      • Ago 2 antibody
      • AGO2_HUMAN antibody
      • Argonaute 2 antibody
      • argonaute 2, RISC catalytic component antibody
      • Argonaute RISC catalytic component 2 antibody
      • Argonaute2 antibody
      • CTA-204B4.6 antibody
      • dAgo2 antibody
      • eIF 2C 2 antibody
      • eIF-2C 2 antibody
      • eIF2C 2 antibody
      • Eif2c2 antibody
      • Eukaryotic translation initiation factor 2C 2 antibody
      • Eukaryotic translation initiation factor 2C subunit 2 antibody
      • hAgo2 antibody
      • MGC3183 antibody
      • PAZ Piwi domain protein antibody
      • PPD antibody
      • Protein argonaute-2 antibody
      • Protein slicer antibody
      • Q10 antibody
      • Slicer protein antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human ovarian carcinoma tissue sections labeling Argonaute -2 with purified ab156870 at 1:100 dilution (1.9 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

    • All lanes : Anti-Argonaute-2 antibody [EPR10410] (ab156870) at 1/5000 dilution (purified)

      Lane 1 : Rat liver lysates
      Lane 2 : Mouse kidney lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 97 kDa

      Blocking and diluting buffer: 5% NFDM/TBST

    • All lanes : Anti-Argonaute-2 antibody [EPR10410] (ab156870) at 1/1000 dilution (purified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : HUVEC (Human umbilical vein endothelial cell) whole cell lysates

      Lysates/proteins at 15 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 97 kDa

      Blocking and diluting buffer: 5% NFDM/TBST

    • Immunocytochemistry/ Immunofluorescence analysis of MCF-7 (Human breast adenocarcinoma epithelial cell) cells labeling Argonaute-2 with Purified ab156870 at 1:200 dilution (9.5μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Ago2 / eIF2C2 with unpurified ab156870 at 1/50 dilution.

    • All lanes : Anti-Argonaute-2 antibody [EPR10410] (ab156870) at 1/1000 dilution (unpurified)

      Lane 1 : HeLa cell lysate
      Lane 2 : MCF7 cell lysate
      Lane 3 : HepG2 cell lysate
      Lane 4 : K562 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat anti-rabbit HRP at 1/2000 dilution

      Predicted band size : 97 kDa
    • Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Argonaute-2 (red) with unpurified ab156870 at a 1/200 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

    • Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling Ago2 / eIF2C2 with unpurified ab156870 at 1/50 dilution.

    References

    This product has been referenced in:
    • Braun J  et al. Rapid identification of regulatory microRNAs by miTRAP (miRNA trapping by RNA in vitro affinity purification). Nucleic Acids Res 42:e66 (2014). WB . Read more (PubMed: 24510096) »
    • Hu Y  et al. Anti-PABPC1 co-immunoprecipitation for examining the miRNAs directly targeting the 3'-UTR of EED mRNA. PLoS One 9:e103695 (2014). WB . Read more (PubMed: 25084349) »

    See all 2 Publications for this product

    Customer reviews and Q&As

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Brain sections)
    Permeabilization
    Yes - 0.2% Tween (added with PBS)
    Specification
    Brain sections
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jul 06 2017

    I am delighted that blocking with BSA rather than milk gave you good results!

    There is no immunogen sequence homology with these protein so this antibody should not detect Ago1, 3, and 4.


    L'anti-Ago2 monoclonal de lapin ab156870

    Human Ago1 : 60%
    Human Ago3 : 60%
    Human Ago4 : 53%



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