Overview

  • Product name
  • Description
    Rabbit polyclonal to Argonaute-2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Drosophila melanogaster
    Does not react with: Human
  • Immunogen

    Synthetic peptide corresponding to Drosophila melanogaster Argonaute-2 aa 550-650 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab24177)

  • Positive control
    • This antibody gave a positive result in WB on Drosophila lysate.
  • General notes
    Please note the immunogen for this antibody has no sequence alignment to human dIFC2. When tested in human, non-specific binding is observed.

Properties

Applications

Our Abpromise guarantee covers the use of ab5072 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
ICC/IF Use at an assay dependent concentration.
WB 1/500 - 1/1000. Detects a band of approximately 55, 95 kDa (predicted molecular weight: 106 kDa).

Detects a band of approximately 55 + 95 kDa (predicted molecular weight: 106 kDa). Swissprot suggests Ago 2 may have 50kDa form. Not yet tested in other applications. Optimal dilutions/concentrations should be determined by the end user.

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

    • Anti-Argonaute-2 antibody (ab5072) at 1 µg/ml + Schneider L2 whole cell lysate (ab14893) at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) at 1/50000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 106 kDa
      Observed band size: 106 kDa
      Additional bands at: 120 kDa, 19 kDa, 50 kDa, 65 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 30 seconds


      This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab5072 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

    • ab5072 staining Ago2 / eIF2C2 (green) in fruit fly (Drosophilia melanogaster) OSS cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.2% Tween and blocked with 1% BSA for 1 hour at 22°C. Samples were incubated with primary antibody (1/200) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated donkey anti-rabbit IgG monoclonal (1/5000) was used as the secondary antibody.

      ab24609 was used to stain Nuclear Pore Complex Proteins (red).

      See Abreview

    • Anti-Argonaute-2 antibody (ab5072) at 1/500 dilution + Drosophila lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab6721)

      Predicted band size: 106 kDa
      Observed band size: 55,95 kDa
      why is the actual band size different from the predicted?



      Rabbit polyclonal to Ago2 (ab5072) at 1/500 on Drosophila lysate (20ug).

      Detects a band of approximately 55 + 95 kDa (predicted molecular weight: 106 kDa). Swissprot suggests Ago 2 may have 50kDa form.

      Secondary antibody - Goat polyclonal to rabbit IgG (HRP) - ab6721.

      Swissprot suggests Ago 2 may have 50kDa form.

    References

    This product has been referenced in:
    • Wang L  et al. Linc-ROR promotes esophageal squamous cell carcinoma progression through the derepression of SOX9. J Exp Clin Cancer Res 36:182 (2017). Read more (PubMed: 29237490) »
    • Zhang J  et al. Circular RNA_LARP4 inhibits cell proliferation and invasion of gastric cancer by sponging miR-424-5p and regulating LATS1 expression. Mol Cancer 16:151 (2017). Read more (PubMed: 28893265) »
    See all 8 Publications for this product

    Customer reviews and Q&As

    1-8 of 8 Abreviews or Q&A

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Brain)
    Permeabilization
    Yes - Tween-20
    Specification
    Brain
    Blocking step
    BSA + Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Jun 21 2017

    Application
    Western blot
    Sample
    Apis mellifera Tissue lysate - whole (Honey bee whole abdomen lysate)
    Gel Running Conditions
    Reduced Denaturing (12% Acrylamide)
    Loading amount
    15 µg
    Treatment
    Infected with recombinant virus Sindbis-GFP
    Specification
    Honey bee whole abdomen lysate
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Michelle Flenniken

    Verified customer

    Submitted Feb 07 2017

    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
    Sample
    Fruit fly (Drosophila melanogaster) Cell (OSS cells)
    Specification
    OSS cells
    Permeabilization
    Yes - Tween 0,2%
    Fixative
    Formaldehyde

    Ms. Nathalie Gred Gueguen

    Verified customer

    Submitted Sep 15 2014

    Answer

    Unfortunately, we do not have any antibodies against Anti-ElF2C2 (hAgo2) which are directly conjugated to a fluorochrome available in our catalog.

    We do however have kits which can be used to conjugate your primary antibody with a fluorochrome. Below are a few kits which may be of interest to you:

    Conjugates for fluorescent detection


    Conjugate
    Color
    3x10μg
    1x100μg
    3x100μg
    1x1mg

    AMCA
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    FITC
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    Rhodamine
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    Cy3.5®
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    B-PE
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    R-PE
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    Cy5®
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    Pe/Cy5®
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    PerCP
    https://www.abcam.com/EasyLink-PerCP-Conjugation-Kit-3-x-10µg-PerCP-ab102908.html https://www.abcam.com/EasyLink-PerCP-Conjugation-Kit-1-x-100µg-PerCP-ab102906.html https://www.abcam.com/EasyLink-PerCP-Conjugation-Kit-3-x-100g-PerCP-ab102907.html https://www.abcam.com/EasyLink-PerCP-Conjugation-Kit-1-x-1mg-PerCP-ab102905.html
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    PE/Cy5.5®
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    APC/Cy5.5®
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    Cy5.5®
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    APC/Cy7®
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    Pleas visit the following webpage for more information regarding these kits: https://www.abcam.com/index.html?pageconfig=resource&rid=13148

    Read More

    Answer

    Thank you very much for your interest in ab5072.

    To our knowledge, ab5072 has not been tested in IP or in a pull down assay.

    We do not use pull down assay as a application on our datasheets. But an antibody that is suitable for IP should also be suitbale for pull down assays.

    Therefore, I can offer a discount off a future purchase if you buy ab5072 now, test it in IP and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of ab5072 off a future order.

    If you are interested in this offer, please follow these steps:

    1. Reply to this e-mail to let me know that you would like to proceed and test ab5072 in IP. I will then send a discount code. This code must be issued before purchasing ab5072 so please wait for my reply before ordering.

    2. Purchase ab5072 either by phone, fax, or online (www.abcam.com).

    3. Test it in IP.

    4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

    5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyproduct ordered and the discount code is valid for 4 months after issue.
    Please remember that submission of the Abreview is sufficient for the discount code to become active.

    We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab5072 turns out to be unsuitable for IP, you will still receive the discount on your next purchase after your Abreview has been submitted.

    Please let me know if you have any questions about this offer and I would be happy to help you further.

    The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

    Read More

    Answer

    Thank you for your enquiry. I sincerely apologize for the error. We just received information regarding the immunogen sequence for this antibody. On investigation by our laboratory, it seems that it does not have any alignment to the human protein. Therefore, this antibody is not expected to react with human samples and therefore, the datasheet has been updated to show the changes. We also received a review from a customer who managed to obtain results with human samples but after confirming the sequence alignment, we deduced that the results obtain by the customer was probably due to non-specific binding. I am unsure about the findings of Schneider MD et al (2006) but our information was taken from SwissProt database. If you have any questions regarding the findings from the authors, I would suggest that you contact them for further details. I am truly sorry that we had a mistake on our datasheet and I am more than happy to offer you a free replacement product of your choice that has the same value as this ab5072, or a refund. Please let me know which method you would prefer and please confirm your purchase details (purchase order number, shipping address/purchasing agent), so that I can immediately issue you the replacement or refund. I look forward to hearing from you again in order to resolve the matter.

    Read More
    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunoprecipitation
    Sample
    Fruit fly (Drosophila melanogaster) Cell lysate - whole cell (Drosophila S2R+ cells)
    Total protein in input
    50 µg
    Specification
    Drosophila S2R+ cells
    Immuno-precipitation step
    Protein A

    Abcam user community

    Verified customer

    Submitted Oct 31 2006

    Answer

    Thank you for your enquiry. Assuming that you are interested in an antibody to control for non-specific binding I would like to recommend that you use an affinity purified rabbit polyclonal antibody against an irrelevant target such as one of the epitope tags. These are obviously foreign to Drosophila and will serve to highlight any non-specificity in your experiments. For example: V5 tag antibody - ChIP Grade (ab15828) This is an immunogen affinity purified sera and supplied as a range: 0.20 - 0.50 mg/ml I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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