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Antibodies

AbFlex® Ago 1/2/3 antibody (rAb)

Catalog No: 91217 Format: 100 µg $375 Buy Now
Catalog No: 91218 Format: 10 µg $75 Buy Now

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Antibody Type:Recombinant
Isotype:IgG2a
Purification:Protein A Chromatography
Host:Mouse
Reactivity:Human

Applications

Immunoprecipitation Validated

for AbFlex Ago 1/2/3 antibody (rAb)Application Notes


Validated Applications:
IP: 5 µg per sample

Application Testing: Parental Monoclonal vs. Recombinant
View PDF file

AbFlex® recombinant antibodies are genetically derived from DNA sequences of parental hybridoma clones. For details on the parental clone, see Catalog No. 39937.

for AbFlex Ago 1/2/3 antibody (rAb)Immunogen

This Ago1/2/3 antibody was raised against a recombinant protein corresponding to amino acids 47-859 of human Ago2.

for AbFlex Ago 1/2/3 antibody (rAb)Buffer

Purified IgG in 140 mM Hepes, pH 7.5, 70 mM NaCl, 32 mM NaOAc, 0.035% sodium azide, 30% glycerol.

for AbFlex Ago 1/2/3 antibody (rAb)References

 
AbFlex<sup>®</sup> Ago 1/2/3 antibody (rAb) tested by immunoprecipitation.

AbFlex Ago1/2/3 antibody (rAb) tested antibody tested by immunoprecipitation.
Active Motif miRNA IP Kit (Cat. No. 25500) was used with 293 cells (10 million cells per IP). Following IP using AbFlex Ago1/2/3 antibody (5 µg per IP) or a negative control IgG, qRT-PCR was performed on samples using primers for IGF2.  Fold enrichment over the negative control IgG shows successful specific IP of IGF2 miRNA associated with the RISC complex. 

for AbFlex Ago 1/2/3 antibody (rAb)Background

AbFlex® antibodies are recombinant antibodies (rAbs) that have been generated using defined DNA sequences to produce highly specific, reproducible antibodies. Each AbFlex antibody contains a 6xHis-Tag, an avidin tag sequence for enzymatic biotin conjugation using the biotin ligase, BirA, and a sortase recognition motif (LPXTG) to attach a variety of labels directly to the antibody including fluorophores, enzymatic substrates (HRP, AP), peptides, drugs as well as solid supports. AbFlex® Ago 1/2/3 antibody was expressed as full-length IgG with mouse immunoglobulin heavy and light chains (IgG2a isotype) in mammalian 293 cells.

The Argonaute family of proteins (Ago1, Ago2, Ago3 and Ago4) are involved in RNAi mediated gene silencing through siRNA and miRNA effectors. The Argonaute proteins are part of RISC, the RNAi Induced Silencing Complex. Binding of RISC can inhibit mRNA translation and results in the degradation of the mRNA through the action of Ago2-mediated target cleavage. Ago1 is non-essential for siRNA-directed target RNA cleavage but is needed for the production of mature miRNA that affects miRNA-directed RNA cleavage. Ago3 lacks RNase activity and does not appear to cleave target mRNA molecules. Argonaute family proteins contain a PAZ and a PIWI domain.

MicroRNA-3´UTR Interaction Products

If you are interested in this antibody, you may also have an interest in products for studying the functional impact of miRNA-3´UTR interactions. The LightSwitch™ 3´UTR Reporter GoClone™ Collection includes over 12,000 human 3´UTRs that have been cloned into the pLightSwitch_3UTR vector for use in luciferase reporter assays. These constructs can be co-transfected with our collections of LightSwitch miRNA Mimics and Inhibitors to study the effects of over-expression of a miRNA of interest, or to knock down the expression of endogenous human miRNAs in living cells.

Bridging Antibody to Improve ChIP, MeDIP and IP

Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use AbFlex® Ago 1/2/3 antibody (rAb).

for AbFlex Ago 1/2/3 antibody (rAb)Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

for AbFlex Ago 1/2/3 antibody (rAb)Guarantee

This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting
 

Technical Data Sheet

AbFlex® Ago 1/2/3 antibody (rAb)
 

Data Thumbnails

AbFlex<sup>®</sup> Ago 1/2/3 antibody (rAb) tested by immunoprecipitation.

Immunoprecipitation of AbFlex® Ago 1/2/3 antibody (rAb).
(Click image to enlarge and see details.)