RIPAb+ CUGBP1 Kit, clone 3B1

• 产品编号：merck-m-03-104      品牌：millipore       原厂货号：03-104
• 产品分类：抗体 > 一抗 > 蛋白特异性一抗
• 应用分类：

描述：

Description:RIPAb+ CUGBP1 - RIP Validated Antibody and Primer Set.

Overview: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.

Background Information: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.

Presentation:Anti-CUGBP1 (Mouse monoclonal IgG). One vial containing 50 μg of protein G purified mouse IgG1 in 50 µL of 0.014 M phosphate buffer, pH 7.6, 0.175 M NaCl, 0.07% sodium azide, and 30% glycerol. Store at -20°C.

Normal Mouse IgG. 
One vial containing 125 μg purified mouse IgG in 125 μL storage buffer containing 0.1% sodium azide. Store at -20°C.

RIP Primers, JUN. One vial containing 75 μL of 5 μM of each primer specific for the cDNA of JUN. Store at -20°C.
FOR: TCG ACA TGG AGT CCC AGG A
REV: GGC GAT TCT CTC CAG CTT CC

Key Applications:

• Western Blotting
• RNA Binding Protein Immunoprecipitation (RIP)
• Immunoprecipitation

原厂资料：

Description:RIPAb+ CUGBP1 - RIP Validated Antibody and Primer Set.

Overview: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.

Background Information: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.

Presentation:Anti-CUGBP1 (Mouse monoclonal IgG). One vial containing 50 μg of protein G purified mouse IgG1 in 50 µL of 0.014 M phosphate buffer, pH 7.6, 0.175 M NaCl, 0.07% sodium azide, and 30% glycerol. Store at -20°C.

Normal Mouse IgG. 
One vial containing 125 μg purified mouse IgG in 125 μL storage buffer containing 0.1% sodium azide. Store at -20°C.

RIP Primers, JUN. One vial containing 75 μL of 5 μM of each primer specific for the cDNA of JUN. Store at -20°C.
FOR: TCG ACA TGG AGT CCC AGG A
REV: GGC GAT TCT CTC CAG CTT CC

Key Applications:

• Western Blotting
• RNA Binding Protein Immunoprecipitation (RIP)
• Immunoprecipitation

注意事项：

Application Notes

RNA Binding Protein Immunoprecipitation:
RIP lysate prepared from HeLa cells (2 X 10⁷ cell equivalents per IP) were subjected to immunoprecipitation using 5 μg of either a normal mouse IgG or Anti-CUGBP1 antibody and the Magna RIP™ Kit (Cat. # 17-700).
Successful immunoprecipitation of CUGBP1-associated RNA was verified by qPCR using primers for ACTB as a negative control and positive control RIP Primers JUN (Please see figures). Data is presented as percent input of each IP sample relative to input total RNA for each amplicon and RIP sample as indicated.
Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.

Immunoprecipitation from RIP lysate:
RIP lysate from HeLa cells (2 X 10⁶ cell equivalents per IP) was subjected to immunoprecipitation using 0.5 μg of either a normal mouse IgG or Anti-CUGBP1 antibody. Precipitated proteins were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-CUGBP1 (1.0 μg/mL). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00231) (Please see figures).

Storage Conditions

Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.