Recombinant Anti-Argonaute-2 antibody [EPR10411] (ab186733)
![Western blot - Anti-Argonaute-2 antibody [EPR10411] (ab186733)](https://www.abcam.com/ps/products/186/ab186733/Images/ab186733-513193-anti-argonaute-2-antibody-epr10411-western-blot-wildtype-hct-116-pools-ago2-knockout-hct-116-u87-mg-mcf7.jpg "Western blot - Anti-Argonaute-2 antibody [EPR10411] (ab186733)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR10411] to Argonaute-2
- Suitable for: Flow Cyt (Intra), ICC/IF, IP, WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Argonaute-2 antibody [EPR10411]
See all Argonaute-2 primary antibodies -
Description
Rabbit monoclonal [EPR10411] to Argonaute-2 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, IP, WB, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- HeLa, MCF7, HepG2, C6 and RAW 264.7 cell lysates; Human cervix carcinoma and Mouse kidney tissues; HeLa and MCF7 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR10411 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab186733 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 |
|---|---|---|
| Flow Cyt (Intra) |
1/60.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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| ICC/IF |
1/100 - 1/250.
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| IP |
1/30 - 1/50.
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| WB |
1/1000 - 1/2000. Detects a band of approximately 97 kDa (predicted molecular weight: 97 kDa).
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| IHC-P |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| Notes |
|---|
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Flow Cyt (Intra)
1/60. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF
1/100 - 1/250. |
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IP
1/30 - 1/50. |
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WB
1/1000 - 1/2000. Detects a band of approximately 97 kDa (predicted molecular weight: 97 kDa). |
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IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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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
modificationsHydroxylated. 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
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Database links
- Entrez Gene: 27161 Human
- Entrez Gene: 239528 Mouse
- Entrez Gene: 59117 Rat
- Omim: 606229 Human
- SwissProt: Q9UKV8 Human
- SwissProt: Q8CJG0 Mouse
- SwissProt: Q9QZ81 Rat
- Unigene: 449415 Human
see all -
Alternative names
- Ago 2 antibody
- AGO2_HUMAN antibody
- Argonaute 2 antibody
see all
Images
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Western blot - Anti-Argonaute-2 antibody [EPR10411] (ab186733)All lanes : Anti-Argonaute-2 antibody [EPR10411] (ab186733) at 1/1000 dilution
Lane 1 : Wild-type HCT 116 cell lysate
Lane 2 : AGO2 knockout HCT 116 cell lysate
Lane 3 : U-87 MG cell lysate
Lane 4 : MCF7 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 97 kDa
Observed band size: 95 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Argonaute-2 antibody [EPR10411] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab186733 was shown to bind specifically to Argonaute-2. A band was observed at 95 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in AGO2 knockout cell line. To generate this image, wild-type and AGO2 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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![Western blot - Anti-Argonaute-2 antibody [EPR10411] (ab186733)](https://www.abcam.com/ps/products/186/ab186733/Images/ab186733-219943-anti-argonaute-2-antibody-epr10411-western-blot.jpg)
Western blot - Anti-Argonaute-2 antibody [EPR10411] (ab186733)All lanes : Anti-Argonaute-2 antibody [EPR10411] (ab186733) at 1/1000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : MCF7 cell lysate
Lane 3 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution
Predicted band size: 97 kDa
Observed band size: 97 kDa -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Argonaute-2 antibody [EPR10411] (ab186733)](https://www.abcam.com/ps/products/186/ab186733/Images/ab186733-219941-anti-argonaute-2-antibody-epr10411-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Argonaute-2 antibody [EPR10411] (ab186733)Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling Ago2 / eIF2C2 with ab186733 at 1/100 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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![Immunocytochemistry/ Immunofluorescence - Anti-Argonaute-2 antibody [EPR10411] (ab186733)](https://www.abcam.com/ps/products/186/ab186733/Images/ab186733-219938-anti-argonaute-2-antibody-epr10411-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Argonaute-2 antibody [EPR10411] (ab186733)Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa cells labeling Ago2 / eIF2C2 with ab186733 at 1/250 dilution followed by Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody at 1/200 dilution. Counter stained with Dapi.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Argonaute-2 antibody [EPR10411] (ab186733)](https://www.abcam.com/ps/products/186/ab186733/Images/ab186733-219940-anti-argonaute-2-antibody-epr10411-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Argonaute-2 antibody [EPR10411] (ab186733)Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Ago2 / eIF2C2 with ab186733 at 1/100 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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![Western blot - Anti-Argonaute-2 antibody [EPR10411] (ab186733)](https://www.abcam.com/ps/products/186/ab186733/Images/ab186733-219942-anti-argonaute-2-antibody-epr10411-western-blot.jpg)
Western blot - Anti-Argonaute-2 antibody [EPR10411] (ab186733)All lanes : Anti-Argonaute-2 antibody [EPR10411] (ab186733) at 1/1000 dilution
Lane 1 : C6 cell lysate
Lane 2 : RAW 264.7 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution
Predicted band size: 97 kDa
Observed band size: 97 kDa -
![Flow Cytometry (Intracellular) - Anti-Argonaute-2 antibody [EPR10411] (ab186733)](https://www.abcam.com/ps/products/186/ab186733/Images/ab186733-219937-anti-argonaute-2-antibody-epr10411-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-Argonaute-2 antibody [EPR10411] (ab186733)Intracellular flow cytometric analysis of2% paraformaldehyde-fixed MCF7 cells labeling Ago2 / eIF2C2 with ab186733 at 1/60 dilution (red) compared to aRabbit monoclonal IgG isotype control (green), followed byGoat anti rabbit IgG (FITC) secondary antibody at 1/150 dilution.
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![Immunoprecipitation - Anti-Argonaute-2 antibody [EPR10411] (ab186733)](https://www.abcam.com/ps/products/186/ab186733/Images/ab186733-219936-anti-argonaute-2-antibody-epr10411-immunoprecipitation.jpg)
Immunoprecipitation - Anti-Argonaute-2 antibody [EPR10411] (ab186733)Western blot analysis of Ago2 / eIF2C2 in MCF7 cell lysate immunoprecipitated with ab186733 at 1/50 dilution.
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution.
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![Anti-Argonaute-2 antibody [EPR10411] (ab186733)](https://www.abcam.com/ps/products/186/ab186733/Images/ab186733-8-benefits-of-recombinant-antibodies.png)
Anti-Argonaute-2 antibody [EPR10411] (ab186733)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (163)
ab186733 has been referenced in 163 publications.
- Liu X et al. LncRNA ZFAS1 contributes to osteosarcoma progression via miR-520b and miR-520e-mediated inhibition of RHOC signaling. Clinics (Sao Paulo) 78:100143 (2023). PubMed: 36473367
- Wang C et al. IGF2BP2-induced circRUNX1 facilitates the growth and metastasis of esophageal squamous cell carcinoma through miR-449b-5p/FOXP3 axis. J Exp Clin Cancer Res 41:347 (2022). PubMed: 36522683
- Chi B et al. Increased expression of miR-194-5p through the circPVRL3/miR-194-5p/SOCS2 axis promotes proliferation and metastasis in pancreatic ductal adenocarcinoma by activating the PI3K/AKT signaling pathway. Cancer Cell Int 22:415 (2022). PubMed: 36539807
- Murmann AE et al. The length of uninterrupted CAG repeats in stem regions of repeat disease associated hairpins determines the amount of short CAG oligonucleotides that are toxic to cells through RNA interference. Cell Death Dis 13:1078 (2022). PubMed: 36585400
- Zhang Y et al. GTF2E2 is a novel biomarker for recurrence after surgery and promotes progression of esophageal squamous cell carcinoma via miR-139-5p/GTF2E2/FUS axis. Oncogene 41:782-796 (2022). PubMed: 34853466