Millipore Sigma Vibrant Logo
Attention: We have moved. EMD Millipore products are no longer available for purchase on emdmillipore.com.Learn More

03-250 RIPAb+™ Ago3 Antibody

Ago3 (Eukaryotic translation initiation factor 2C 3) belongs to the argonaute family & contains a PAZ & a Piwi domain. Ago3 plays a role in the RNA interference (RNAi) pathway.

RIPAb+™ Ago3 Antibody MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Key Applications: Immunocytochemistry, Immunoprecipitation, RNA Binding Protein Immunoprecipitation (RIP) Research Category: Epigenetics & Nuclear Function Research Sub-Category: RNA Binding Protein (RBP)
View Products on Sigmaaldrich.com
03-250
10 assays  10 assays per set. Recommended use: ~5 µg of antibody per RIP (dependent upon biological context).
Purchase on Sigma-Aldrich

Special Offers

Overview

Replacement Information

Special Offers

100% Performance Guaranteed

Key Specifications Table

Species ReactivityKey Applications
HICC, IP, RIP
Description
Catalogue Number03-250
Trade Name
  • RIPAb+
  • Upstate
DescriptionRIPAb+™ Ago3 Antibody
OverviewRIPAb+ antibodies are evaluated using the RNA Binding Protein Immunoprecipitation (RIP) assay. Each RIPAb+ antibody set includes a negative control antibody to ensure specificity of the RIP reaction and is verified for the co-immunoprecipitation of RNA associated specifically with the immunoprecipitated RNA-binding protein of interest. Where appropriate, the RIPAb+ set also includes quantitative RT-PCR control primers (RIP Primers) to biologically validate your IP results by successfully co-precipitating the specific RNA targets, such as messenger RNAs. The qPCR protocol and primer sequences are provided, allowing researchers to validate RIP protocols when using the antibody in their experimental context. If a target specific assay is not provided, the RIPAb+ kit is validated using an automated microfluidics-based assay by enrichment of detectable RNA over control immunoprecipitation.
Ago3 (Eukaryotic translation initiation factor 2C 3) belongs to the argonaute family and contains a PAZ and a Piwi domain. Ago3 plays a role in the RNA interference (RNAi) pathway, which is important for heterochromatin formation and accurate chromosome segregation.
Alternate Names
  • eIF-2C 3
  • eIF2C 3
  • FLJ12765
  • MGC86946
Background InformationEukaryotic translation initiation factor 2C 3, Ago3, belongs to the argonaute family and contains 1 PAZ and 1 Piwi domain.
Ago3 is indicated to play a role in the RNA interference (RNAi) pathway, which is important for heterochromatin formation and accurate chromosome segregation. Unlike Ago1 and Ago2, Ago3 and AG04 do not possess the ability to dissociate miRNA duplexes in a minimal in vitro system. Re-introduction of any Ago variant into embyronic (ES) cells deficient for expression of all four Ago variants will rescue miRNA silencing defects and reduce apoptosis.
References
Product Information
FormatPurified
Control
  • Includes normal rabbit IgG and primers specific for human IGF2 mRNA.
PresentationAnti-Ago3 (Mouse Monoclonal). One vial containing 50 µg of protein G purified monoclonal in buffer containing 0.1 M Tris-glycine (pH 7.4), 150mM NaCl, 0.05% sodium azide before the addition of glycerol to 30%. Store at -20° C.
Concentration: 0.7 mg/mL
Normal mouse IgG. One vial containing 125 µg purified mouse IgG in 125 µL storage buffer containing 0.1% sodium azide. Store at -20°C.
RIP Primer IGF2. One vial containing 75 μL of 5 μM of each primer specific for human IGF2 mRNA. Store at
-20°C.
FOR: GCG GCT TCT ACT TCA GCA G
REV: CAG GTG TCA TAT TGG AAG AAC
Quality LevelMQ100
Applications
ApplicationAgo3 (Eukaryotic translation initiation factor 2C 3) belongs to the argonaute family & contains a PAZ & a Piwi domain. Ago3 plays a role in the RNA interference (RNAi) pathway.
Key Applications
  • Immunocytochemistry
  • Immunoprecipitation
  • RNA Binding Protein Immunoprecipitation (RIP)
Application NotesRNA Binding Protein Immunoprecipitation (microRNA):

Representative lot data.
RIP Lysate prepared from 293 cells (2 X 107 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal mouse IgG (Cat. # CS200621), or Anti-Ago 3 antibody (Cat. # CS207366) and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of Ago 3-associated microRNA was verified by TaqMan® microRNA assay, hsa-miR-15a. (Figure2). Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.
Immunocytochemistry Analysis:
Representative lot data.
Confocal fluorescent analysis of HeLa cells using Anti-Ago3, clone 4B1-F6 (Red). Actin filaments have been labeled with Alexa Fluor® 488 dye-Phalloidin (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
(Alexa Fluor is a registered trademark of Life Technologies®.)
Immunoprecipitation Analysis: A representative lot was used by an independent laboratory in HeLa cells. (Azuma-Mukai, A., et al. (2008). Proc Natl Acad Sci USA. 105(23):7964-7969.)
Biological Information
ImmunogenRecombinant protein corresponding to human Ago3
Clone4B1-F6
ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
HostMouse
Specificity This antibody has been shown to not cross react with Ago1, Ago2, or Ago4 (Azuma-Mukai A et al. (2008) Proc. Natl. Acad. Sci. USA 105: 7964-7969).
IsotypeIgG2aκ
Species Reactivity
  • Human
Antibody TypeMonoclonal Antibody
Entrez Gene Number
Entrez Gene SummaryThis gene encodes a member of the Argonaute family of proteins which play a role in RNA interference. The encoded protein is highly basic, contains a PAZ domain and a PIWI domain, and may play a role in short-interfering-RNA-mediated gene silencing. This gene is located on chromosome 1 in a tandem cluster of closely related family members including argonaute 4 and eukaryotic translation initiation factor 2C, 1. Two transcript variants encoding distinct isoforms have been identified for this gene.
Gene Symbol
  • EIF2C3
  • AGO3
Purification MethodProtein G Purified
UniProt Number
UniProt SummaryFUNCTION: Required for RNA-mediated gene silencing (RNAi). Binds to short RNAs such as microRNAs (miRNAs) and represses the translation of mRNAs which are complementary to them. Lacks endonuclease activity and does not appear to cleave target mRNAs.

SUBUNIT STRUCTURE: Interacts with EIF4B, IMP8, PRMT5 and TNRC6B.

SUBCELLULAR LOCATION: Cytoplasm > P-body.
Molecular Weight~96 kDa
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality AssuranceRNA Binding Protein Immunoprecipitation:
Representative lot data
RIP Lysate prepared from 293 cells (2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal mouse IgG , or 5 µg Anti-Ago 3 antibody and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of Argonaute 3-associated RNA was verified by qPCR using RIP Primers IGF2, (Figure 1).
Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.
Usage Statement
  • 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.
Storage and Shipping Information
Storage ConditionsStable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
Packaging Information
Material Size10 assays
Material Package10 assays per set. Recommended use: ~5 µg of antibody per RIP (dependent upon biological context).
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalog Number GTIN
03-250 04053252439421

Related Products & Applications

Related Products

Catalog Number Description
MABE144 Anti-Ago3 Antibody, clone 4B1-F6

Related Products By: Brand Facete

RIPAb+™

Categories

Life Science Research > Antibodies and Assays > Immunoassays > Immunoprecipitation (IP) > RNA-Binding Protein Immunoprecipitation (RIP) > RIP Validated Antibodies
Life Science Research > Antibodies and Assays > Primary Antibodies
Life Science Research > Protein Detection and Quantification > Immunoassays > Immunoprecipitation (IP) > RNA-Binding Protein Immunoprecipitation (RIP) > RIP Validated Antibodies