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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| B, H, M, R, Ca | RIP, WB, IP |
| Description | |
|---|---|
| Catalogue Number | 03-191 |
| Brand Family | Upstate |
| Trade Name |
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| Description | RIPAb+ Upf1 - 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. Upf1 is part of a post-splicing multiprotein complex involved in both mRNA nuclear export and mRNA surveillance. Upf1 is located only in the cytoplasm. When translation ends, it interacts with the protein that is a functional homolog of yeast Upf2p to trigger mRNA decapping. |
| Alternate Names |
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| Background Information | Upf1 is part of a post-splicing multiprotein complex involved in both mRNA nuclear export and mRNA surveillance. mRNA surveillance detects exported mRNAs with truncated open reading frames and initiates nonsense-mediated mRNA decay (NMD). When translation ends upstream from the last exon-exon junction, this triggers NMD to degrade mRNAs containing premature stop codons. Upf1 is located only in the cytoplasm. When translation ends, it interacts with the protein that is a functional homolog of yeast Upf2p to trigger mRNA decapping. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
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| Presentation | Anti-Upf1 (Rabbit Polyclonal). One vial containing 70 μL of purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide and 30% glycerol. Store at -20°C. Normal Rabbit IgG. One vial containing 125 µg of rabbit IgG in 125 µL of storage buffer containing 0.05% sodium azide. Store at -20°C. RIP Primers, ARF1. One vial containing 75 μL of 5 μM of each primer specific for human ARF1. Store at -20°C. FOR: GAC CAC GAT CCT CTA CAA GC REV: TCC CAC ACA GTG AAG CTG ATG |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This RIPAb+ Upf1 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. |
| Key Applications |
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| Application Notes | Immunoprecipitation from RIP lysate: Representative lot data. RIP lysate from HeLa cells (~2 X 10E7 cell equivalents per IP) was subjected to immunoprecipitation using either 7 µL of a normal rabbit IgG or 7 µL of Anti-Upf1 antibody. Ten percent of the precipitated proteins (lane 1: rabbit IgG, lane 2: Upf1) were resolved by electrophoresis, transferred to PVDF and probed with anti-Upf1 antibody (1:2000). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00232) Arrow indicates Upf1 (~125 kDa) (Please see figures). Western Blot Analysis: Representative lot data. U2OS cell lysate was resolved by electrophoresis, transferred to PVDF membrane and probed with anti-Upf1 (0.5 μg/mL). Proteins were visualized using a donkey anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates Upf1 (~125 kDa) (Please see figures). |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated synthetic linear peptide corresponding to Upf1. |
| Host | Rabbit |
| Specificity | This antibody detects Upf1. |
| Species Reactivity |
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| Species Reactivity Note | Demonstrated to react with human. Predicted to react with mouse, rat, bovine and canine based on sequence homology. |
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | This gene encodes a protein that is part of a post-splicing multiprotein complex involved in both mRNA nuclear export and mRNA surveillance. mRNA surveillance detects exported mRNAs with truncated open reading frames and initiates nonsense-mediated mRNA decay (NMD). When translation ends upstream from the last exon-exon junction, this triggers NMD to degrade mRNAs containing premature stop codons. This protein is located only in the cytoplasm. When translation ends, it interacts with the protein that is a functional homolog of yeast Upf2p to trigger mRNA decapping. Use of multiple polyadenylation sites has been noted for this gene. [provided by RefSeq]. |
| Gene Symbol |
|
| Purification Method | Affinity Purfied |
| UniProt Number |
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| UniProt Summary | FUNCTION: Part of a post-splicing multiprotein complex. Involved in nonsense-mediated decay (NMD) as part of the SMG1C complex, a mRNA surveillance complex that recognizes and degrades mRNAs containing premature translation termination codons (PTCs). The complex probably acts by associating with ribosomes during tranlation termination on mRNPs. If an exon junction complex (EJC) is located 50-55 or more nucleotides downstream from the termination codon, RENT1 is phosphorylated by SMG1, triggering nonsense-mediated decay (NMD). Essential for embryonic viability. SUBUNIT STRUCTURE: Found in a complex with RENT1, RENT2, RENT3A and RENT3B. Found in a complex with PARN. Found in a post-splicing complex with SMG1, NXF1, RBM8A, RENT1, RENT2, RENT3A, RENT3B and RNPS1. Found in a mRNA decay complex with EXOSC2, EXOSC4, EXOSC10, PARN, XRN1, DCP2, RENT1, RENT2 and RENT3B. Interacts with EST1A and RENT2. Component of the SMG1C complex, at least composed of SMG1, SMG8 and SMG9. The SMG1C complex is then recruited on premature translation termination codons (PTCs) to form the ribosome:SURF complex, at least composed of ERF1, ERF3 (ERF3A or ERF3B), EEF2, UPF1/RENT1, SMG1, SMG8 and SMG9. Interacts (when hyperphosphorylated) with PNRC2. SUBCELLULAR LOCATION: Cytoplasm. Cytoplasm › P-body. Note: Hyperphosphorylated form is targeted to the P-body, while unphosphorylated protein is distributed throughout the cytoplasm. TISSUE SPECIFICITY: Ubiquitous. DOMAIN: The [ST]-Q motif constitutes a recognition sequence for kinases from the PI3/PI4-kinase family. PTM: Phosphorylated by SMG1; required for formation of mRNA surveillance complexes. Phosphorylated upon DNA damage, probably by ATM or ATR. SEQUENCE SIMILARITIES: Belongs to the DNA2/NAM7 helicase family. Contains 1 C2H2-type zinc finger. |
| Molecular Weight | ~125 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | RNA Binding Protein Immunoprecipitation: RIP Lysate prepared from HeLa cells (2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using either 7 µL of a normal rabbit IgG or 7 µL Anti-Upf1 antibody and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700). Successful immunoprecipitation of Upf1-associated RNA was verified by qPCR using RIP Primers ARF1 (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 |
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| 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: ~7 μL of antibody per RIP (dependent upon biological context). |