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03-113
Sigma-AldrichRIPAb+ p54nrb/NonO - RIP Validated Antibody and Primer Set
Each RIPAb+ antibody set includes a negative control antibody to ensure specificity of the RIP reaction & is verified for the co-IP of RNA associated specifically with the immunoprecipitated RNA binding protein of interest.
More>>Each RIPAb+ antibody set includes a negative control antibody to ensure specificity of the RIP reaction & is verified for the co-IP of RNA associated specifically with the immunoprecipitated RNA binding protein of interest. Less<<
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RIPAb+ p54nrb/NonO - RIP Validated Antibody and Primer Set
Overview
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.
Alternate Names
Non-POU domain-containing octamer-binding protein
NonO protein
54 kDa nuclear RNA- and DNA-binding protein
NMT55
Background Information
p54nrb/NonO is a DNA and RNA binding protein, that binds the conventional octamer sequence in double stranded DNA, as well as single-stranded DNA and RNA at a site independent of the duplex site. P54nrb/NonO is also involved in pre-mRNA splicing.
References
Product Information
Format
Purified
Control
Includes negative control mouse IgG antibody and control primers specific for human FOS.
Presentation
Anti-p54nrb/NonO (Mouse Monoclonal IgG). One vial containing 50 µg of protein G purified mouse monoclonal IgG in 50 µL of buffer containing (0.1 M Tris-glycine, 0.15 M NaCl, pH 7.4, 0.05% 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, FOS. One vial containing 75 μL of 5 μM of each primer specific for human FOS. Store at -20°C. FOR: GAG AGC TGG TAG TTA GTA GCA TGT TGA REV: AAT TCC AAT AAT GAA CCC AAT AGA TTA GTT A
Each RIPAb+ antibody set includes a negative control antibody to ensure specificity of the RIP reaction & is verified for the co-IP of RNA associated specifically with the immunoprecipitated RNA binding protein of interest.
Key Applications
RNA Binding Protein Immunoprecipitation (RIP)
Immunoprecipitation
Western Blotting
Immunohistochemistry
Application Notes
Immunoprecipitation from RIP lysate: Representative lot data. RIP lysate from HeLa cells (2 X 106 cell equivalents per IP) was subjected to immunoprecipitation using either 0.5 µg of a normal mouse IgG or 0.5 µg of Anti-p54nrb/NONO antibody. Precipitated proteins (lane 1: rabbit IgG, lane 2: anti-p54nrb/NONO) and HeLa whole cell lysate (lane 3) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-p54nrb/NONO antibody (0.5 µg/mL). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00231) (Please see figures).
Western Blot Analysis: 0.5-2.0 μg/mL of a previous lot detected p54nrb/NonO in RIPA lysates from MCF-7 cells (Please see figures).
Immunohistochemistry: This antibody has been reported by an independent laboratory to detect p54nrb/NonO in human breast tumor sections.
Immunoprecipitation: Reported by an independent lab to immunoprecipitate p54nrb/NonO.
Biological Information
Immunogen
KLH-conjugated, synthetic peptide corres-ponding to amino acids TVREAGPPAFYRPNS
This gene encodes an RNA-binding protein which plays various roles in the nucleus, including transcriptional regulation and RNA splicing. A rearrangement between this gene and the transcription factor E3 gene has been observed in papillary renal cell carcinoma. Alternatively spliced transcript variants have been described. Pseudogenes exist on Chromosomes 2 and 16.
Function: DNA- and RNA binding protein, involved in several nuclear processes. Binds the conventional octamer sequence in double stranded DNA. Also binds single-stranded DNA and RNA at a site independent of the duplex site. Involved in pre-mRNA splicing, probably as an heterodimer with SFPQ. Interacts with U5 snRNA, probably by binding to a purine-rich sequence located on the 3' side of U5 snRNA stem 1b. The SFPQ-NONO heteromer associated with MATR3 may play a role in nuclear retention of defective RNAs. The SFPQ-NONO heteromer may be involved in DNA unwinding by modulating the function of topoisomerase I/TOP1. The SFPQ-NONO heteromer may be involved in DNA nonhomologous end joining (NHEJ) required for double-strand break repair and V(D)J recombination and may stabilize paired DNA ends. In vitro, the complex strongly stimulates DNA end joining, binds directly to the DNA substrates and cooperates with the Ku70/G22P1-Ku80/XRCC5 (Ku) dimer to establish a functional preligation complex. Nono is involved in transcriptional regulation. The SFPQ-NONO-NR5A1/SF-1 complex binds to the CYP17 promoter and regulates basal and cAMP-dependent transcriptional avtivity. NONO binds to an enhancer element in long terminal repeats of endogenous intracisternal A particles (IAPs) and activates transcription. Subunit structure: Monomer and component of the SFPQ-NONO complex, which is probably a heterotetramer of two 52 kDa (NONO) and two 100 kDa (SFPQ) subunits. NONO is a component of spliceosome and U5.4/6 snRNP complexes. Interacts with PSPC1 and SNRPA/U1A. Part of complex consisting of SFPQ, NONO and MATR3. Part of a complex consisting of SFPQ, NONO and NR5A1/SF-1. Part of a complex consisting of SFPQ, NONO and TOP1. Interacts with SPI1 By similarity. Interacts with RNF43. Subcellular location: Nucleus. Tissue specificity: Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas. Also found in a number of breast tumor cell lines. Post-translational modification: The N-terminus is blocked. Involvement in disease: A chromosomal aberration involving NONO may be a cause of papillary renal cell carcinoma (PRCC). Translocation t(X;X)(p11.2;q13.1) with TFE3. Sequence similarities: Contains 2 RRM (RNA recognition motif) domains.
Molecular Weight
54 kDa
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
RNA Binding Protein Immunoprecipitation: RIP Lysate prepared from HeLa cells (2 X 107 cell equivalents per IP) were subjected to immunoprecipitation using either 5 µg of a normal mouse IgG or 5 µg of Anti-p54nrb/NONO antibody and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700). Successful immunoprecipitation of p54nrb/NONO-associated RNA was verified by qPCR using RIP Primers FOS (Please see figures). Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.
Usage Statement
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.
Storage and Shipping Information
Storage Conditions
Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon receipt, 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. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
Packaging Information
Material Size
10 assays
Material Package
10 assays per set. Recommended use: ~5 μg of antibody per RIP (dependent upon biological context).
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Bestellnummer
GTIN
03-113
04053252679759
Documentation
RIPAb+ p54nrb/NonO - RIP Validated Antibody and Primer Set SDB
