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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | WB, IP, ICC, IHC | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MABE56 |
| Description | Anti-pan Ago Antibody, clone 2A8 |
| Alternate Names |
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| Background Information | Argonaute (Ago) proteins are a family of proteins of ~95 kDa that bind directly to mature miRNAs. In mammals, there are four Ago proteins (Ago1-4). This family of proteins is a central component of most mammalian miRNPs that mediates important functions in small RNA-directed regulatory pathways. The Ago family consists of two subclases, Ago and Piwi containing two characteristic domains known as PAZ and PIWI. These two domains interact with different parts of the mature miRNA. The PAZ domain interacts with the 3’-end of the miRNA, and the PIWI domain interacts with the 5’ phosphate and 5’-proximal region of the miRNA to guide target mRNA recognition. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
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| Presentation | Purified mouse monoclonal IgG1κ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-pan Ago Antibody, clone 2A8 is a Mouse Monoclonal Antibody for detection of pan Ago also known as protein argonaute-2 or PAZ Piwi domain protein & has been validated in WB, IP, ICC & IHC. |
| Key Applications |
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| Application Notes | Immunoprecipitation Analysis: A representative lot was used by an independent laboratory in IP. (Nelson, P., et al. (2007). RNA. 13:1787–1792.) Immunocytochemistry Analysis: A representative lot was used by an independent laboratory in IC. (Nelson, P., et al. (2007). RNA. 13:1787–1792.) Immunohistochemistry Analysis: A representative lot was used by an independent laboratory in IH. (Nelson, P., et al. (2007). RNA. 13:1787–1792.) |
| Biological Information | |
|---|---|
| Immunogen | Histidine - tagged recombinant protein corresponding to human Ago. |
| Clone | 2A8 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| Isotype | IgG1κ |
| Species Reactivity |
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| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | This gene encodes a member of the Argonaute family of proteins which play a role in RNA interference. The encoded protein is highly basic, and contains a PAZ domain and a PIWI domain. It may interact with dicer1 and play a role in short-interfering-RNA-mediated gene silencing. Multiple transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq]. |
| Gene Symbol |
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| Purification Method | Protein G |
| UniProt Number |
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| UniProt Summary | 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. CATALYTIC ACTIVITY: Endonucleolytic cleavage to 5-phosphomonoester. ENZYME REGULATION: Inhibited by EDTA. SUBUNIT STRUCTURE: Interacts with DICER1 through its Piwi domain and with TARBP2 during assembly of the RNA-induced silencing complex (RISC). Together, DICER1, EIF2C2/AGO2 and TARBP2 constitute the trimeric RISC loading complex (RLC), or micro-RNA (miRNA) loading complex (miRLC). Within the RLC/miRLC, DICER1 and TARBP2 are required to process precursor miRNAs (pre-miRNAs) to mature miRNAs and then load them onto EIF2C2/AGO2. EIF2C2/AGO2 bound to the mature miRNA constitutes the minimal RISC and may subsequently dissociate from DICER1 and TARBP2. Note however that the term RISC has also been used to describe the trimeric RLC/miRLC. The formation of RISC complexes containing siRNAs rather than miRNAs appears to occur independently of DICER1. Interacts with EIF2C1/AGO1. Also interacts with DDB1, DDX6, DDX20, DDX47, DHX9, DHX36, EIF6, FXR1, GEMIN4, ILF3, MOV10, PRMT5, P4HA1, P4HB, SART3, TNRC6A, TNRC6B and UPF1. Interacts with the P-body components DCP1A and XRN1. SUBCELLULAR LOCATION: Cytoplasm › P-body. Nucleus. Note: Translational repression of mRNAs results in their recruitment to P-bodies. Translocation to the nucleus requires IMP8. 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). PTM: Hydroxylated. 4-hydroxylation appears to enhance protein stability but is not required for miRNA-binding or endonuclease activity. SEQUENCE SIMILARITIES: Belongs to the argonaute family. Ago subfamily. Contains 1 PAZ domain. Contains 1 Piwi domain. BIOPHYSIOCHEMICAL PROPERTIES: Kinetic parameters: KM=1.1 nM for a synthetic 21-nucleotide single-stranded RNA Ref.7 Ref.18 SEQUENCE CAUTION: The sequence AAH07633.1 differs from that shown. Reason: Erroneous initiation. The sequence AAL76093.1 differs from that shown. Reason: cDNA contains a duplication of an internal sequence at the 5 end. The sequence BC125214 differs from that shown. Reason: Frameshift at position 450. |
| Molecular Weight | ~ 95 and 70 kDa observed. This antibody recognizes Ago1, Ago2, Ago3, and Ago4. Note that this antibody has been reported to cross-react with radixin (~70 kDa). (Nelson, P., et al. (2007). RNA. 13:1787–1792.) |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blot in HeLa cell lysate. Western Blot Analysis: 0.5 µg/mL of this antibody detected Ago on 10 µg of HeLa cell lysate. |
| Usage Statement |
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| Storage and Shipping Information | |
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| Storage Conditions | Stable for 1 year at 2-8°C from date of receipt. |
| Packaging Information | |
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| Material Size | 100 µg |