Overview

  • Product name
    Anti-Argonaute-2 antibody - ChIP Grade
    See all Argonaute-2 primary antibodies
  • Description
    Rabbit polyclonal to Argonaute-2 - ChIP Grade
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IP, IHC-P, ChIP, RIP, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Rabbit, CowDoes not react with: Drosophila melanogaster
  • Immunogen

    Synthetic peptide corresponding to Argonaute-2 aa 350-450 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab32380)

  • Positive control
    • WB: HeLa whole cell lysate. IHC-P: Human breast cancer tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab32381 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
IP Use at an assay dependent concentration. PubMed: 24280866
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ChIP Use at an assay dependent concentration. PubMed: 19696410
RIP Use at an assay dependent concentration.
WB Use a concentration of 2 µg/ml. Detects a band of approximately 87 kDa (predicted molecular weight: 97 kDa).
ICC/IF 1/200.

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

  • Immunohistochemical analysis of Argonaute2 (AGO2) in human tissues sections from slowly progressive, rapidly progressive, or normal biopsies.

    Representative images of slowly progressive (A–B, E–F), rapidly progressive (C–D, G–H), and normal (I–J) biopsies stained with IgG control (A, C, E, G, & I) and anti-AGO2 antibody (B, D, F, H, & J) are shown. Images A, B, C, & D were stained with antibody concentrations of 20 µg/ml and images E, F, G, H, I, & J were stained with an antibody concentration of 4 µg/ml. Sections were counterstained with hematoxylin. Protein expression stains brown in this procedure (original magnification: ×200).

    Panels A-D shown.

  • All lanes : Anti-Argonaute-2 antibody - ChIP Grade (ab32381) at 2 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat whole cell lysate (ab7899)
    Lane 3 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 97 kDa
    Observed band size: 87 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 55 kDa. We are unsure as to the identity of these extra bands.



    The identification of the 55 kDa band is unclear but this band has also been observed in HeLa lysates in the Western Blot of ab5072, targeting Drosophila Ago2 / eIF2C2.

  • ab32381 (1/200) staining Ago2/ eIF2C2 in asynchronous HeLa (Human epithelial cell line from cervix adenocarcinoma) cells (green). Cells were fixed with paraformaldehyde, permeabilized with 0.5% TritonX100/ PBS and counterstained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.

    See Abreview

  • IHC image of Ago2/eIF2C2 staining in human breast cancer FFPE section, performed on a BondTM system using the standard protocol F.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32381, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX.

  • RIP analysis of 368T1 murine lung cancer cells using ab32381 to bind Ago2 / eIF2C2.

    Cells were transfected with an empty vector (empty), a vector expressing wild-type Hmga2 (wt), a vector expressing a let-7 site mutated Hmga2 (m7), a vector expressing wild-type Hmga2 without a start codon (ATG wt), or a vector expressing a let-7 site mutated Hmga2 without a start codon (ATG m7). RIP was performed with 5 µg Ago2 / eIF2C2 or IgG antibody per 107 cells. Binding was detected using qRT-PCR.

    See Abreview

References

This product has been referenced in:
  • Liang W  et al. Knockdown of growth-arrest specific transcript 5 restores oxidized low-density lipoprotein-induced impaired autophagy flux via upregulating miR-26a in human endothelial cells. Eur J Pharmacol 843:154-161 (2019). Read more (PubMed: 30468731) »
  • Dong H  et al. Activation of LncRNA TINCR by H3K27 acetylation promotes Trastuzumab resistance and epithelial-mesenchymal transition by targeting MicroRNA-125b in breast Cancer. Mol Cancer 18:3 (2019). Read more (PubMed: 30621694) »
See all 79 Publications for this product

Customer reviews and Q&As

1-10 of 10 Q&A

Question
Answer

We do not have any cross-reactivity data for this product with other argonaute proteins. However, the sequence homology analysis suggests it is unlikely this antibody would cross-react with Ago1, Ago3 or Ago4. Human Ago1 : 69% homology Human Ago3 : 54% homology Human Ago4 : 46% homology

Read More

Answer

Thank you for contacting us.No, we have not stained mouse primary cells for confocal microscopy, although mouse tissue has been tested as well as human cell cultures.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for your reply.

I have sent you 2 new vials of ab32381, since the original 2 we sent did not work in western blotting for you. Your new order number is********and you should hopefully have them by Friday.

If there is anything else I can help you with, please let me know.

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Question
Answer

Thank you for contacting Abcam earlier today.

I just noticed on that we shipped 2 vials of ab32381 on the order information that you provided. I just wanted to check if both of these vials were for your lab and therefore need both vials replaced. Or if one of the vials was for another lab.

As soon as I hear back from you, I will send the replacements.

Looking forward to your reply.

Read More

Question
Answer

Thank you for contacting Abcam.

The alignment with mouse is also excellent with a 98% homology. Staining of mouse tissue using mouse antibody is a complicated process as high levels of background are often observed. It is notoriously difficult to eliminate this background. This particular application is not covered under our abpromise.

Please let me know if you have any questions.

Read More

Answer

Thank you very much for your interest in ab32381 and ab57113.

Bothantibodie arenot tested and guaranteed for rat and CLIP. It is tested for mouse samples but also not for CLIP. We do not know yet if it will work. Since rat is predicted to workwith ab32381 I suggest to try this antibody.


Therefore, I can offer a discount off a future purchase if you buy ab32381 now, test it inCLIP with mouse samplesand 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 1 free primary antibody.

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 ab32381 in CLIP with mouse samples. I will then send a discount code. This code must be issued before purchasing ab32381 so please wait for my reply before ordering.

2. Purchase ab32381 either by phone, fax, or online (www.abcam.com).

3. Test it in CLIP with mouse samples.

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 anyprimary antibodyordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab32381 turns out to be unsuitable for CLIP with mouse samples, 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

Question
Answer

Thank you for contacting Abcam.

The alignment with rat is also excellent with a 95% homology. Again this is not covered under our abpromise but is eligible for the Abreview discount program

Please let me know if you have any questions.

Read More

Answer

Thank you for contacting us.

We do offer another product, ab57113, which has been tested in ChIP and has a better signal. This product has not yet been tested in mouse however. I ran an alignment between the human Ago2 sequence used for creating this product and mouse Ago2. The results were excellent with 95% homology. As such, I believe that you should have no problems at all using this product in mouse but because this has not been tested in mouse, it would not be covered under our Abpromise guarantee.

Should you be interested this would qualify for our Abreview 100% testing discount. Under this program you purchase ab57113 to test it on mouse, You then review the performance of the product by creating an Abreview on our site. After you have created the Abreview, it takes about 5-10 minutes, you would be entitled to 1 free antibody of your choice.

Please let me know if you are interested or have any other questions.

Read More

Answer

Thank you for contacting us. The alignment results I obtained are: SpAgo1 and SpAgo2 69% identity PmAgo1/2 75% identity. There are some gaps, rather than contiguous conserved sequence, and I am reluctant to recommend this antibody for detection of your proteins. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

At this time we have no data regarding the use of ab32381 in CLIP. So we would not be able to offer the Abpromise for the use of this antibody in CLIP. I would happy to set up a discount code for your customer to test ab32381 in CLIP.

Read More

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