Ago2/eIF2C2 Antibody (2A8)

Western Blot: Ago2/eIF2C2 Antibody (2A8) [NBP2-59159] - Western blot analysis using the antibodyWhole cell extracts (40 ug) from HeLa cells were analysed by Western blot using the antibody diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (kDa) is shown on the left.

Immunocytochemistry/Immunofluorescence: Ago2/eIF2C2 Antibody (2A8) [NBP2-59159] - HeLa cells were stained with the antibody against Ago and with DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labelled with the Ago antibody (middle) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa Fluor 594. The left panel shows staining of the nuclei with DAPI. A merge of both stainings is shown on the right.

Immunoprecipitation: Ago2/eIF2C2 Antibody (2A8) [NBP2-59159] - IP of endogenous Argonaute proteins using the Ago antibody. HeLa cells (10 cm dish per IP) were subjected to UV crosslinking, lysed in 70 uL of 1X PXL buffer, incubated on ice for 10 min, then treated with DNase I. Dynabeads Protein A (40 uL each) were incubated with 12 ug anti-mouse IgG, used as a bridge antibody, briefly washed, and then incubated with antibody (4 or 12 ug), or non-immune mouse IgG control (4 or 12 ug). The cell lysate and Dynabeads were mixed and incubated at 4C O/N. The beads were washed twice at 4C for 5 min using 500 uL of (i) 1X PXL buffer, (ii) 5X PXL (5X PBS, 0.1% SDS, 0.5% Na-deoxycholate, 0.5% NP-40), and (iii) 1X PNK (50 mM Tris pH7.4, 10 mM MgCl2, 0.5% NP-40). 10 uL of input (10% of starting material), 10 uL of supernatants after IP (10%), and all of immunoprecipitants (~90%) were loaded on SDS-PAGE gel and subjected to Western blot analysis using the antibody and the ECL plus reagent.

Immunoprecipitation: Ago2/eIF2C2 Antibody (2A8) [NBP2-59159] - Allelic imbalance Ago-RIP assay with the antibody to show the preferential co-immunoprecipitation of miRNA targeted alleles. HeLa cells were transfected with: pRL-TK-4A + pRL-TK-4G + pcDNA-miR-1 (miR-1 is supposed to target Aandallele) pRL-TK-4A + pRL-TK-4G + pcDNA-miR-1C (miR-1C is supposed to target Gandallele) pRL-TK-4A + pRL-TK-4G + pcDNA-miR-377 (miR-377 is supposed to target neither allele). Cell lysate was immunoprecipitated with Protein G agarose beads incubated with antibody. RNA was isolated from the complexes from the input and IP samples, reverse transcribed using random hexamers, and subjected to RT-PCR to confirm enrichment of miRNA targeted allele in IP sample compared to input sample. PCR products were directly sequenced. The electropherograms at the polymorphic site are shown in the second panel. The predicted miRNA target alleles were efficiently co-immunoprecipitated by the antibody.

• Reactivity: Hu, Mu
• Applications: WB, ICC/IF, IHC, IP
• Clone: 2A8
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

Ago2/eIF2C2 Antibody (2A8) Summary

• Immunogen: The exact immunogen is propietary information.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: AGO2
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/Immunofluorescence 1:500
  • Immunohistochemistry 1:500
  • Immunoprecipitation 8 ug/IP
  • Western Blot 1:500

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Purity: Protein A purified

Alternate Names for Ago2/eIF2C2 Antibody (2A8)

• AGO2argonaute 2
• argonaute2
• EC 3.1.26.n1
• EC 3.1.26.n2
• eIF2C 2
• eIF-2C 2
• Eukaryotic translation initiation factor 2C 2
• eukaryotic translation initiation factor 2C, 2
• hAgo2
• MGC3183
• PAZ Piwi domain protein
• PPD
• protein argonaute-2
• Protein slicer
• Q10

Background

Ago2 / eIF2C2 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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Ago2/eIF2C2 Antibody (NBP2-59159)(1)

Publication using NBP2-59159

• Kim S, Ha J, Choi S et al. Use of Bacterial Extracellular Vesicles for Gene Delivery to Host Cells Biomolecules 2022-08-24 [PMID: 36139009] (IP, E. coli)

Bioinformatics

• Gene Symbol: AGO2