Overview

  • Product name
    Anti-Argonaute-2 antibody [2E12-1C9] - ChIP Grade
    See all Argonaute-2 primary antibodies
  • Description
    Mouse monoclonal [2E12-1C9] to Argonaute-2 - ChIP Grade
  • Host species
    Mouse
  • Tested applications
    Suitable for: CHIPseq, WB, ICC/IF, IHC-P, IP, Flow Cyt, ChIPmore details
  • Species reactivity
    Reacts with: Chicken, Human, Zebrafish
    Does not react with: Xenopus laevis
  • Immunogen

    Recombinant fragment corresponding to Human Argonaute-2 aa 483-859.

  • General notes

    This product was changed from ascites to tissue culture supernatant on 30th April 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab57113 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
CHIPseq Use at an assay dependent concentration. PubMed: 23592263
WB Use at an assay dependent concentration. Predicted molecular weight: 97 kDa.
ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
IP Use at an assay dependent concentration. PubMed: 19574298
Flow Cyt Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

ChIP Use at an assay dependent concentration. PubMed: 20010811

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

  • All lanes : Anti-Argonaute-2 antibody [2E12-1C9] - ChIP Grade (ab57113) at 5 µg/ml

    Lane 1 : Argonaute-2 overexpressing HEK-293 cells line
    Lane 2 : Argunaiute-2 overexpressing cell line transfected with Argonaute-2 RNAi.

    Predicted band size: 97 kDa



    The loading control is GAPDH.

    This image was generated using the ascites version of the product.

  • Western Blot detection against the recombinant fragment imunogen (68.4 KDa).

    This image was generated using the ascites version of the product.

  • Ago2 / eIF2C2 antibody (ab57113) used in immunofluorescence at 10ug/ml on HeLa cells.

    This image was generated using the ascites version of the product.

  • Ago2 / eIF2C2 antibody (ab57113) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human stomach.

    This image was generated using the ascites version of the product.

  • Overlay histogram showing HeLa cells stained with ab57113 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab57113, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.

    This image was generated using the ascites version of the product.

References

This product has been referenced in:
  • Bu JY  et al. Long non-coding RNA LINC00978 promotes cell proliferation and tumorigenesis via regulating microRNA-497/NTRK3 axis in gastric cancer. Int J Biol Macromol 123:1106-1114 (2019). Read more (PubMed: 30452981) »
  • Bellissimo T  et al. Argonaute 2 drives miR-145-5p-dependent gene expression program in breast cancer cells. Cell Death Dis 10:17 (2019). Read more (PubMed: 30622242) »
See all 93 Publications for this product

Customer reviews and Q&As

1-10 of 31 Abreviews or Q&A

Application
Western blot
Sample
Mouse Serum (mouse plasma)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
mouse plasma
Blocking step
gelatin as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Apr 01 2019

Application
Immunoprecipitation
Immuno-precipitation step
Protein G
Sample
Human Cell lysate - other (Cytoplasmic extract, A549 cells)
Specification
Cytoplasmic extract, A549 cells
Total protein in input
1000 µg

Abcam user community

Verified customer

Submitted Feb 19 2015

This product is known to not work in this application or species.
Application
Western blot
Loading amount
20 cells
Gel Running Conditions
Reduced Denaturing
Sample
Xenopus laevis Cell lysate - whole cell (oocytes)
Specification
oocytes
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Nov 21 2013

Answer

I can confirm that after checking with some colleagues, the cross reactivity of ab57113 with rabbit IgG has not been tested yet. Therefore, we have no related data to share with you at this moment.

Read More

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
https://www.abcam.com/EasyLink-FITC-Conjugation-Kit-3-x-10µg-FITC-ab102885.html - https://www.abcam.com/EasyLink-FITC-Conjugation-Kit-3-x-100µg-FITC-ab102884.html https://www.abcam.com/EasyLink-FITC-Conjugation-Kit-1-x-1mg-FITC-ab102883.html
Cy3®
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Rhodamine
https://www.abcam.com/EasyLink-Rhodamine-Conjugation-Kit-3-x-10µg-Rhodamine-ab102915.html - https://www.abcam.com/EasyLink-Rhodamine-Conjugation-Kit-3-x-100µg-Rhodamine-ab102914.html https://www.abcam.com/EasyLink-Rhodamine-Conjugation-Kit-1-x-1mg-Rhodamine-ab102913.html
Cy3.5®
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B-PE
- https://www.abcam.com/EasyLink-B-Phycoerythrin-Conjugation-Kit-1-x-100µg-B-PE-ab102870.html https://www.abcam.com/EasyLink-B-Phycoerythrin-Conjugation-Kit-3-x-100µg-B-PE-ab102871.html https://www.abcam.com/EasyLink-B-Phycoerythrin-Conjugation-Kit-1-x-1mg-B-PE-ab102869.html
R-PE
https://www.abcam.com/EasyLink-R-Phycoerythrin-Conjugation-Kit-3-x-10g-R-PE-ab102919.html https://www.abcam.com/EasyLink-R-Phycoerythrin-Conjugation-Kit-1-x-100-µg-R-PE-ab102917.html https://www.abcam.com/EasyLink-R-Phycoerythrin-Conjugation-Kit-3-x-100µg-R-PE-ab102918.html https://www.abcam.com/EasyLink-R-Phycoerythrin-Conjugation-Kit-1-x-1mg-R-PE-ab102916.html
Texas Red®
https://www.abcam.com/EasyLink-Texas-Red-Conjugation-Kit-3-x-10µg-TR-ab102925.html - https://www.abcam.com/EasyLink-Texas-Red-Conjugation-Kit-3-x-100µg-TR-ab102924.html https://www.abcam.com/EasyLink-Texas-Red-Conjugation-Kit-1-x-1mg-TR-ab102923.html
Cy5®
https://www.abcam.com/EasyLink-Cy5-Conjugation-Kit-3-x-10µg-Cy5-ab102877.html - https://www.abcam.com/EasyLink-Cy5-Conjugation-Kit-3-x-100µg-Cy5-ab102876.html https://www.abcam.com/EasyLink-Cy5-Conjugation-Kit-1-x-1mg-Cy5-ab102875.html
Pe/Cy5®
https://www.abcam.com/EasyLink-PE-Cy5-Conjugation-Kit-3-x-10µg-PE-Cy5-ab102896.html https://www.abcam.com/EasyLink-PE-Cy5-Conjugation-Kit-1-x-100µg-PE-Cy5-ab102894.html https://www.abcam.com/EasyLink-PE-Cy5-Conjugation-Kit-3-x-100µg-PE-Cy5-ab102895.html https://www.abcam.com/EasyLink-PECy5-Conjugation-Kit-1-x-1mg-PECy5-ab102893.html
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
PerCP/Cy5.5®
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PE/Cy5.5®
https://www.abcam.com/EasyLink-PE-Cy5-5-Conjugation-Kit-3-x-10µg-PE-Cy5-5-ab102900.html https://www.abcam.com/EasyLink-PE-Cy5-5-Conjugation-Kit-1-x-100µg-PE-Cy5-5-ab102898.html https://www.abcam.com/EasyLink-PE-Cy5-5-Conjugation-Kit-3-x-100µg-PE-Cy5-5-ab102899.html https://www.abcam.com/EasyLink-PECy55regnbspConjugation-Kit-1-x-1mg-PECy55reg-ab102897.html
APC/Cy5.5®
https://www.abcam.com/EasyLink-APC-Cy5-5-Conjugation-Kit-3x10µg-APC-Cy5-5-ab102855.html https://www.abcam.com/EasyLink-APC-Cy5-5-Conjugation-Kit-1x100µg-APC-Cy5-5-ab102853.html https://www.abcam.com/EasyLink-APC-Cy5-5-Conjugation-Kit-3x100µg-APC-Cy5-5-ab102854.html https://www.abcam.com/EasyLink-APC-Cy5-5-Conjugation-Kit-1-x-1mg-APC-Cy5-5-ab102852.html
Cy5.5®
- - https://www.abcam.com/EasyLink-Cy5-5-Conjugation-Kit-3-x-100µg-Cy5-5-ab102879.html https://www.abcam.com/EasyLink-Cy5-5-Conjugation-Kit-1-x-1mg-Cy5-5-ab102878.html
APC/Cy7®
https://www.abcam.com/EasyLink-APC-Cy7-Conjugation-Kit-3-x-10µg-APC-Cy7-ab102859.html https://www.abcam.com/EasyLink-APC-Cy7-Conjugation-Kit-1x100µg-APC-Cy7-ab102857.html https://www.abcam.com/EasyLink-APC-Cy7-Conjugation-Kit-3x100µg-APC-Cy7-ab102858.html https://www.abcam.com/EasyLink-APC-Cy7-Conjugation-Kit-1-x-1mg-APC-Cy7-ab102856.html
PE/Cy7®
https://www.abcam.com/EasyLink-PECy7-Conjugation-Kit-3-x-10g-PECy7-ab102904.html https://www.abcam.com/EasyLink-PE-Cy7-Conjugation-Kit-1-x-100µg-PE-Cy7-ab102902.html https://www.abcam.com/EasyLink-PE-Cy7-Conjugation-Kit-3-x-100µg-PE-Cy7-ab102903.html https://www.abcam.com/EasyLink-PE-Cy7-Conjugation-Kit-1-x-1mg-PE-Cy7-ab102901.html
Cy7®
https://www.abcam.com/EasyLink-Cy7-Conjugation-Kit-3-x-10g-Cy7-ab102882.html - https://www.abcam.com/EasyLink-Cy7-Conjugation-Kit-3-x-100µg-Cy7-ab102881.html https://www.abcam.com/EasyLink-Cy7-Conjugation-Kit-1-x-1mg-Cy7-ab102880.html



Pleas visit the following webpage for more information regarding these kits: https://www.abcam.com/index.html?pageconfig=resource&rid=13148

Read More

Answer

1.
Our data for ab57113 on zebrafish samples comes from an abreview submitted by a customer on 23 March 2011 showing positive results in western blot. These results can be found at the following webpage:
https://www.abcam.com/Ago2--eIF2C2-antibody-ChIP-Grade-ab57113-reviews.html?intabreviewid=23495

Our data for ab57113 on chicken samples comes from a publication which used ab57113 in ChIP in chicken. See the following publication:
Giles KE et al. Maintenance of a constitutive heterochromatin domain in vertebrates by a Dicer-dependent mechanism. Nat Cell Biol 12:94-9; sup pp 1-6 (2010).

2)
The image of the western blot on the datasheet is detecting not the full length Ago2 protein but the recombinant fragment imunogen corresponding to amino acids 483-859 of Human Ago2 / eIF2C2 (UniProt: Q9UKV8). This recombinant fragment is therefore 68.4 KDa and this is the band detected on the western blot.

We do have data to show that a band at ˜95 kDa is seen in western blots:

1. https://www.abcam.com/Ago2--eIF2C2-antibody-ChIP-Grade-ab57113-reviews.html?intabreviewid=23111.
2. Behrman S et al. A CHOP-regulated microRNA controls rhodopsin expression. J Cell Biol 192:919-27 (2011).
3. Joyce CE et al. Deep sequencing of small RNAs from human skin reveals major alterations in the psoriasis miRNAome. Hum Mol Genet 20:4025-40 (2011).
4. Wang X et al. Moloney leukemia virus 10 (MOV10) protein inhibits retrovirus replication. J Biol Chem 285:14346-55 (2010).
5. Li X et al. Identification of sequence features that predict competition potency of siRNAs. Biochem Biophys Res Commun 398:92-7 (2010).

Read More

Answer

Thank you for contacting us in regards to the Anti-Ago2 / eIF2C2 antibody - ChIP Grade (ab57113) and I am sorry for the delay in getting back to you.

I now have further information to share with you. The anti-Ago2 / eIF2C2 antibody - ChIP Grade (ab57113) is described as being raised against the "recombinant full length protein, corresponding to amino acids 1-377 of Human Ago2 / eIF2C2". However, the sequence that this refers to is the NCBI Genebank protein entry AAH07633.1. This corresponds to the region 483-859 of the Uniprot entry Q9UKV8.

I am sorry for the confusion this has caused. This would explain why the antibody has not been able to detect the truncation you have been expressing.


I hope this has been of some help. I look forward to receiving your reply.

Read More

Answer

Thank you for your email.

I can confirm that the 6 units of ab57113 were added to your order. You will get 5 units of lot GB90870-1 from the UK, one unit with lot GB90870-1 will be requested from the US.

I hope you will find everything after your liking. Please do not hesitate to contact us again with further questions.

Read More

Answer

Thank you for contacting me again.

At the moment we have five units of ab57113 with the same lot. The currently available lot number is GR90870-1. If you like to order the product you should send your order as soon as possible and include the lot number in your order.

I hope I was able to help you giving you this information and wish you a nice evening.

Read More
Application
Immunocytochemistry
Sample
Human Cultured Cells (HELA)
Specification
HELA
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton-X in PBS
Blocking step
H2O2 as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Oct 23 2012

1-10 of 31 Abreviews or Q&A

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