RIPAb+ Ago2; clone 9E8.2

Stock Code: 3586254
Manufacturer Part No: 03-110
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Analysis Note


ControlIncludes negative control mouse IgG antibody and control primers specific for human FOS.


Application


RNA Binding Protein Immunoprecipitation (microRNA):
RIP Lysate prepared from HeLa cells (2 X 107 cell equivalents per IP) were subjected to immunoprecipitation using 5 µL of either a normal mouse IgG or 5 µL of Anti-Ago2 antibody and the Magna RIP® RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of Ago2-associated microRNA was verified by TaqMan™ microRNA assay, hsa-miR-23a. (Please see figures).
Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.


Immunoprecipitation from RIP lysate:
Representative lot data.
RIP lysate from HeLa cells (2 X 106 cell equivalents per IP) was subjected to immunoprecipitation using 0.5 µL of either a normal mouse IgG or Anti-Ago2 antibody. Precipitated proteins (lane 1: mouse IgG, lane 2: anti-Ago2) and HeLa whole cell lysate (lane 3) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Ago2 antibody (1:5,000 dilution).
Proteins were visualized using goat anti-mouse secondary antibody conjugated to HRP and chemiluminescence detection.
(Please see figures).


Research Sub CategoryRNA Metabolism & Binding ProteinsRNA Binding Protein (RBP)


Research CategoryEpigenetics & Nuclear Function


This RIPAb+ Ago2 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.


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


Ago2 (argonaute-2), also known as eukaryotic translation initiation factor 2C (EIF2C2), is an essential component for siRNA-directed RNA interference (RNAi) response. Ago2 is an endonuclease required for the unwinding of siRNA duplex and assembly of siRNA into RNA-induced silencing complexes (RISC). Ago2 interacts with DICER1 through its Piwi domain. This Piwi domain is thought to provide RNA cleavage activity via a mechanism similar to RNase H. Ago2 activity is necessary for embryonic development as well as RNA-mediated gene silencing (RNAi).


Immunogen


KLH-conjugated, synthetic peptide containing the sequence YSGAGPALAPPAPPPPIQG.


Legal Information


UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany


MAGNA RIP is a registered trademark of Merck KGaA, Darmstadt, Germany


Packaging


10 assays per set. Recommended use: ~5 µL of antibody per RIP (dependent upon biological context).


Physical form


Anti-Ago2 (Mouse Monoclonal IgG1κ). One vial containing 50 µL of ascites with 0.05% sodium azide. Store at -20°C.

Normal Mouse IgG

Quality Level100, 500
ManufacturerSIGMA-ALDRICH

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