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

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1-10 of 10 Abreviews

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

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

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Baboon Tissue sections (uterus)
Specification
uterus
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: antigen unmasking solution
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jul 05 2012

Application
CLIP
Sample
Human Cell lysate - whole cell (HEK 293T)
Specification
HEK 293T
Type
iCLIP
Irradiation Energy Level (mJ/cm2): 150
Wavelength (nm): 254
Positive control
Anti-hnRNP C antibody (sc-15386) bound protein G dynabeads, 0.1 mg/ml (antibody/beads)
Negative control
Protein G dynabeads without antibody bound
Immuno-precipitation step
Other - Protein G dynabeads

Abcam user community

Verified customer

Submitted Mar 21 2012

This product is known to not work in this application or species.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Xenopus laevis Cell (egg extract with sperm nuclei)
Specification
egg extract with sperm nuclei
Fixative
Formaldehyde
Permeabilization
Yes - methanol
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 17 2011

This product is known to not work in this application or species.
Application
Western blot
Sample
Xenopus laevis Cell lysate - other (egg extract)
Loading amount
50 µg
Specification
egg extract
Gel Running Conditions
Non-reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 17 2011

Application
Western blot
Sample
Zebrafish Tissue lysate - whole (whole 4dpf embryos)
Loading amount
60 µg
Specification
whole 4dpf embryos
Gel Running Conditions
Reduced Denaturing (precast 4-15%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Viviana Anelli

Verified customer

Submitted Mar 23 2011

Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa, HEK293)
Loading amount
40 µg
Specification
HeLa, HEK293
Gel Running Conditions
Reduced Denaturing (7.5%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Feb 16 2011

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