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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, M | WB, IP | Rb | Affinity Purified | Polyclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 07-1016 |
| Description | Anti-phospho-Upf1 (Ser1127) Antibody |
| Alternate Names |
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| Background Information | Regulator of nonsense transcripts 1 (UniProt: Q92900; also known as ATP-dependent helicase RENT1, Nonsense mRNA reducing factor 1, NORF1, Up-frameshift suppressor 1 homolog, hUpf1) is encoded by the UPF1 (also known as KIAA0221, RENT1) gene (Gene ID: 5976) in human. Upf1 is a member of the DNA2/NAM7 helicase family. It is a RNA-dependent helicase and ATPase required for nonsense-mediated decay (NMD) of mRNAs containing premature stop codons. It is recruited to mRNAs upon translation termination and undergoes a cycle of phosphorylation and dephosphorylation. It is phosphorylated by SMG1 and phosphorylation is considered as an essential step in NMD and is required for formation of mRNA surveillance complexes. Phosphorylated Upf1 is recognized by EST1B/SMG5, SMG6, and SMG7, which provide a link to the mRNA degradation machinery involving exonucleolytic and endonucleolytic pathways. A hyper-phosphorylated form is targeted to the P-body, while unphosphorylated protein is distributed throughout the cytoplasm. The ATPase activity of Upf1 is required for disassembly of mRNPs undergoing NMD and is also essential for embryonic viability. Two isoforms of Upf1 have been reported that are produced by alternative splicing. Upf1 contains a UPF1-type zinc-finger domain (aa 121-272) and a nucleotide-binding domain (aa 506-510). |
| Product Information | |
|---|---|
| Format | Affinity Purified |
| Control |
|
| Presentation | Purified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-phospho-Upf1 (Ser1127), Cat. No. 07-1016, is a highly specific rabbit polyclonal antibody that targets Regulator of nonsense transcripts 1 (RENT1) phosphorylated at Ser1127 and has been tested in Immunoprecipitation, peptide Inhibition Assay, and Western Blotting. |
| Key Applications |
|
| Application Notes | Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected phospho-Upf1 (Ser1127) in 10 µg of A431 treated with Calyculin A and Okadaic Acid. Peptide Inhibition Analysis: A 1:1,000 dilution from a representative lot blocked phospho-Upf1 (Ser1127) in NIH3T3 treated with Calyculin A and Okadaic Acid. Immunoprecipitation Analysis: 5 µg from a representative lot immunoprecipitated phospho-Upf1 (Ser1127) in NIH3T3 treated with Calyculin A and Okadaic Acid. |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated linear peptide corresponding to Upf1 phosphorylated at Ser1127. |
| Epitope | Phosphorylated Ser1127 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Rabbit |
| Specificity | This rabbit polyclonal antibody detects Regulator of nonsense transcripts 1 (RENT1) in multiple species. It targets an epitope within 9 amino acids surrounding phosphorylated Ser1127. |
| Species Reactivity |
|
| Species Reactivity Note | Human, Mouse. Predicted to react with Yeast, Chicken, Zebra Fish, Bovine, Rat based on 100% sequence homology. |
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
|
| 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 |
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| Modifications |
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| Purification Method | Affinity Purfied |
| UniProt Number |
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| UniProt Summary | FUNCTION: Plays a role in replication-dependent histone mRNA degradation at the end of phase S. 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 UPF1, UPF2, UPF3A and UPF3B. Found in a complex with PARN. Found in a post-splicing complex with SMG1, NXF1, RBM8A, UPF1, UPF2, UPF3A, UPF3B and RNPS1. Found in a mRNA decay complex with EXOSC2, EXOSC4, EXOSC10, PARN, XRN1, DCP2, UPF1, UPF2 and UPF3B. Interacts with EST1A, UPF2 and SLBP. 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. Ref.16 Ref.8 Ref.11 Ref.15 Ref.24 SUBCELLULAR LOCATION: Cytoplasm. Cytoplasm › P-body. Note: Hyperphosphorylated form is targeted to the P-body, while unphosphorylated protein is distributed throughout the cytoplasm. TISSUE SPECIFICTY: 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. SEQUENCE CAUTION: The sequence BAA19664.2 differs from that shown. Reason: Erroneous initiation. |
| Molecular Weight | ~140 kDa observed. 125 kDa calculated. |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blotting in NIH3T3 treated with Calyculin A and Okadaic Acid. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected phospho-Upf1 (Ser1127) in 10 µg lysate from NIH3T3 treated with Calyculin A (50 nM) and Okadaic Acid (500 nM) for 30 minutes. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at 2-8°C from date of receipt. |
| Packaging Information | |
|---|---|
| Material Size | 100 µg |