Anti-Argonaute-2抗体- ChIP Grade (ab32381)

概述

  • 产品名称Anti-Argonaute-2抗体- ChIP Grade
    参阅全部 Argonaute-2 一抗
  • 描述
    兔多克隆抗体to Argonaute-2 - ChIP Grade
  • 经测试应用适用于: IP, IHC-P, ChIP, RIP, WB, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
    预测可用于: Rabbit, Cow不与反应: Drosophila melanogaster
  • 免疫原

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

  • 阳性对照
    • ab32381 gives a positive result in Western Blot in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line), Jurkat (Human T cell lymphoblast-like cell line), HeLa (Human epithelial carcinoma cell line) Nuclear Lysate

性能

应用

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.

应用 Ab评论 说明
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.

靶标

  • 功能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.
  • 序列相似性Belongs to the argonaute family. Ago subfamily.
    Contains 1 PAZ domain.
    Contains 1 Piwi domain.
  • 结构域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).
  • 翻译后修饰Hydroxylated. 4-hydroxylation appears to enhance protein stability but is not required for miRNA-binding or endonuclease activity.
  • 细胞定位Cytoplasm > P-body. Nucleus. Translational repression of mRNAs results in their recruitment to P-bodies. Translocation to the nucleus requires IMP8.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

Anti-Argonaute-2 antibody - ChIP Grade 图像

  • 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
    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 following publication; http://www.ibiosci.or.kr/home/data/2004/0080213.pdf and in the Western Blot of ab5072, targeting Drosophila Ago2 / eIF2C2.

  • 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 haematoxylin and mounted with DPX.
  • ab32381 (1/200) staining Ago2/ eIF2C2 in assynchrnous HeLa cells (green). Cells were fixed with paraformaldehyde, permeabilised 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

  • 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

Anti-Argonaute-2 antibody - ChIP Grade (ab32381)参考文献

This product has been referenced in:
  • Yin Y  et al. Interleukin 7 up-regulates CD95 protein on CD4+ T cells by affecting mRNA alternative splicing: priming for a synergistic effect on HIV-1 reservoir maintenance. J Biol Chem 290:35-45 (2015). WB ; Human . Read more (PubMed: 25411246) »
  • Scherr M  et al. Differential expression of miR-17~92 identifies BCL2 as a therapeutic target in BCR-ABL-positive B-lineage acute lymphoblastic leukemia. Leukemia 28:554-65 (2014). IP . Read more (PubMed: 24280866) »

See all 18 Publications for this product

Product Wall

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 : ...

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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 co...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application RIP
Sample Mouse Cell lysate - whole cell (murine lung cancer cells)
Specification murine lung cancer cells
Type RIP without cross-linking
Duration of cross-linking step: 0 second(s)
Detection step Real-time PCR
Positive control empty vector cells for miR-10a
Negative control RIP with control IgG
Immuno-precipitation step Protein G
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Abcam user community

Verified customer

提交于 Nov 15 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.5 % Tritionx100
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
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Abcam user community

Verified customer

提交于 Oct 05 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (Mouse nueroblastoma N2A cell)
Loading amount 30 µg
Specification Mouse nueroblastoma N2A cell
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Abcam user community

Verified customer

提交于 Jul 03 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (saos-2 human osteosarcoma cell line)
Specification saos-2 human osteosarcoma cell line
Fixative Paraformaldehyde
Permeabilization Yes - PBS TRITON 0.25%
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
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Abcam user community

Verified customer

提交于 Apr 19 2012

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 anyth...

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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 repla...

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Abcam has not validated the combination of species/application used in this Abreview.
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 A dynabeads, 0.1 mg/ml (antibody/beads)
Negative control Protein A dynabeads without antibody bound
Immuno-precipitation step Other - Protein A dynabeads
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Abcam user community

Verified customer

提交于 Mar 21 2012

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...

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