Myotonic dystrophy (MD) is an autosomal dominant neuromuscular disease that is associated with a (CTG)n repeat expansion in the 3′-untranslated region of the myotonin protein kinase (Mt-PK) gene. A (CUG)n oligonucleotides triplet repeat pre-mRNA/mRNA binding protein may play an important role in DM pathogenesis. HeLa cell protein, CUG-BP1, has been purified based upon its ability to bind specifically to (CUG)8 oligonucleotides in vitro. CUG-BP1 is the major (CUG)8 binding activity in normal cells. CUG-BP1 has been identified as isoforms of a novel heterogeneous nuclear ribonucleoprotein (hnRNP), hNab50. The CUG-BP/hNab50 protein is localized predominantly in the nucleus and is associated with polyadenylated RNAs in vivo. In vitro RNA-binding/photocrosslinking studies demonstrate that CUG-BP/hNab50 binds to RNAs containing the Mt-PK 3′-UTR.

RIPAb+ antibodies are evaluated using the RNA Binding Protein Immunoprecipitation (RIP) assay. Each RIPAb+ antibody set includes quantitative RT-PCR control primers (RIP Primers) to biologically validate your IP results by successfully coprecipitating a specific RNA target (where such a specific target is known). The qPCR protocol and primer sequences are provided, allowing researchers to validate RIP protocols when using the antibody in their experimental context. Each set also includes a negative control antibody to ensure specificity of the RIP reaction.

The RIPAb+ CUGBP1 set includes the CUGBP1 (CUG triplet repeat, RNA binding protein 1) antibody, a negative control antibody (normal mouse IgG), and positive qPCR primers which amplify a 100 bp fragment of the human cDNA of JUN. The CUGBP1 and negative control antibodies are supplied in a scalable "per RIP" reaction size and can be used to functionally validate the precipitation of CUGBP1-associated RNA.

The CUGBP1 antibody is made against full-length GST fusion protein corresponding to human CUGBP1, also known as CUG triplet repeat, RNA binding protein 1 or heterogeneous nuclear ribonucleoprotein (hnRNP) hNab50.

This RIPAb+ CUGBP1 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

RNA Binding Protein Immunoprecipitation:
RIP lysate prepared from HeLa cells (2 X 10⁷ cell

RNA Binding Protein Immunoprecipitation:<br />RIP lysate prepared from HeLa cells (2 x 10⁷cell equivalents per IP) were subjected to immunoprecipitation using 5 ug of either a normal mouse IgG, or Anti-CUGBP1 antibody and the Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of CUGBP1-associated RNA was verified by qPCR using RIP Primers JUN.
Please refer to the Magna RIP (Cat. # 17-700) or EZ-Magna RIP (Cat. # 17-701) protocol for experimental details.

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