RIPAb+ AUF1

Stock Code: 3586255
Manufacturer Part No: 03-111

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Descriptions

Analysis Note

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

Application

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 µg of either a normal rabbit IgG or Anti-AUF1 antibody. Precipitated proteins (Lane 1: rabbit IgG, Lane 2: anti-AUF1) and HeLa whole cell lysate (Lane 3) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-AUF1 antibody (1.0 µg/mL).
Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00231) (Please see figures).

Western Blot Analysis: 0.01-1 µg/mL of a previous lot detected AUF1 in RIPA lysates from HeLa nuclear extract and A431 cells.

This RIPAb+ AUF1 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

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

Research CategoryEpigenetics & Nuclear Function

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

RIPAb+ 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.

A+U Rich RNA Binding Factor (AUF1) is comprised of four isoforms of 37, 40, 42, and 45 kDa. Although the role of each isoform has yet to be fully characterized, a direct correlation has been observed between each AUF1 isoform's binding affinity and its RNA destabilizing activity toward different AREs (A + U rich element in the 3' untranslated region), with the isoforms p37 and p42 being the most effective.
AUF1 is a bcl2 mRNA binding protein and potentially all its isoforms are able to form complexes with the bcl2 ARE. ARE mediated bcl2 mRNA downregulation during apoptosis involves AUF1 and suggest different roles for its four isoforms.

Immunogen

Human AUF1 purified by Ni²⁺ affinity column.

Legal Information

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