Anti-pan Ago Antibody, clone 2A8 ZooMAb® Mouse Monoclonal
SIGMA/ZMS1071 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZMS1071, Anti-pan Ago, clone 2A8 ZooMAb® Mouse Monoclonal. Reactivity was detected using a Rabbit Anti-Mouse IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in tubules and glomeruli of human kidney tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZMS1071, Anti-pan Ago, clone 2A8 ZooMAb® Mouse Monoclonal. Reactivity was detected using a Rabbit Anti-Mouse IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in tubules and glomeruli of human kidney tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZMS1071, Anti-pan Ago, clone 2A8 ZooMAb® Mouse Monoclonal. Reactivity was detected using a Rabbit Anti-Mouse IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in tubules and glomeruli of human kidney tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of MCF-7 cells was performed using a 1:100 dilution of Cat. No. ZMS1071, Anti-pan Ago, clone 2A8 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of MCF-7 cells was performed using a 1:100 dilution of Cat. No. ZMS1071, Anti-pan Ago, clone 2A8 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of MCF-7 cells was performed using a 1:100 dilution of Cat. No. ZMS1071, Anti-pan Ago, clone 2A8 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from HeLa cells (Lane 1) and MCF-7 cells (Lane 2) were probed with Cat. No. ZMS1071, Anti-pan Ago, clone 2A8 ZooMAb® Mouse Monoclonal (1:1,000 dilution). Proteins were visualized using a Goat Anti-Mouse IgG1 secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates Ago protein isoforms 1 and 2 (~95 kDa and 70 kDa).
Flow Cytometry Enhanced Validation - Recombinant Antibody Technology Staining of one million MCF-7 cells was performed using 1 μg of Cat. No. ZMS1071, Anti-pan Ago, clone 2A8 ZooMAb® Mouse Monoclonal (Yellow histogram) or the equivalent amount of a Mouse IgG1 isotype control (Grey histogram), followed by a PE-conjugated Goat Anti-MouseIgG1 secondary antibody.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | mouse |
| clone | 2A8, monoclonal |
| recombinant monoclonal | |
| conjugate | unconjugated |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG1κ |
| mol wt | calculated mol wt ~97.21 and 93.62 kDa |
| observed mol wt ~95 and 70 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| storage temp. | 2-8°C |
| technique(s) | flow cytometry: suitable |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| immunoprecipitation (IP): suitable | |
| western blot: suitable | |
| UniProt accession no. | Q9UKV8  |
| Application: | Quality Control Testing Evaluated by Western Blotting in HeLa cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected pan Ago in HeLa cell lysate. Tested Applications Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected pan Ago in MCF-7 cell lysate. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected pan Ago in human kidney tissue sections. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected pan Ago in MCF-7 cells. Flow Cytometry Analysis: 1 μg from a representative lot detected pan Ago in one million MCF-7 cells. Immunoprecipitation Analysis: A representative lot immunoprecipitated Ago protein in Immunoprecipitation applications (Nelson, P.T., et. al. (2007). RNA. 13(10):1787-92). Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Application: | Anti-pan Ago, clone 2A8 ZooMAb®, Cat. No. ZMS1071, is a recombinant Mouse monoclonal antibody that targets Ago protein and is used in Flow Cytometry, Immunocytochemistry, Immunohistochemistry (Paraffin), Immunoprecipitation, and Western Blotting. |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb antibodies are reliably available and ready to ship when you need them. |
| Immunogen: | His-tagged recombinant fragment corresponding to 813 amino acids from human Argonaute-2 protein. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant mouse monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 2A8 is a ZooMAb® mouse recombinant monoclonal antibody that detects all isoforms of human Argonaute-2 protein. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | Protein argonaute-2 (UniProt: Q9UKV8; also known as Argonaute2, hAgo2, Argonaute RISC catalytic component 2, Eukaryotic translation initiation factor 2C, eIF-2C, eIF2C, PAZ Piwi domain protein, PPD, Protein slicer) is encoded by the AGO2 (also known as EIF2C2) gene (Gene ID: 27161) in human. Ago2 is a highly conserved member of the AGO family of proteins with catalytic activity and is required for RNA-mediated gene silencing (RNAi) by the RNA-induced silencing complex (RISC). Binding of RISC to a perfectly complementary mRNA generally results in silencing due to endonucleolytic cleavage of the mRNA specifically by Ago2. Binding of RISC to a partially complementary mRNA results in silencing through inhibition of translation and is independent of endonuclease activity. Ago2 is reported to inhibit translation initiation by binding to the 7-methylguanosine cap, thereby preventing the recruitment of the translation initiation factor eIF4-E. 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. Ago2 is shown to have four core domains that are known as N domain, PAZ domain (aa 235-348), MID domain, and PIWI domain (aa 517-818). The PAZ domain is an RNA binding module that recognizes the 3 end of both siRNA and miRNA, in a sequence independent manner and is reported to be strictly required for Ago2 to form the RISC complex. The PIWI domain has major roles within Ago2, including RNase H like endonucleolytic activity and regulating the interaction between MID domain and DNA/RNA. Ago2 is shown to undergo 4-hydoxylation that enhances its stability but is not essential for miRNA binding or endonuclease activity. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Ye, Z., et al. (2015). J. Cancer 6(9); 877-882). |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |