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07-1016 Sigma-AldrichAnti-phospho-Upf1 (Ser1127) Antibody

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.

Anti-phospho-Upf1 (Ser1127) Antibody MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

SDS
CoA
References

Research Category: Epigenetics & Nuclear Function Research Sub-Category: RNA Metabolism & Binding Proteins Gene Symbol: UPF1, RENT1 UniProt Number: Q92900

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Key Spec 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	Regulator of nonsense transcripts 1 ATP-dependent helicase RENT1 Nonsense mRNA reducing factor 1 NORF1 Up-frameshift suppressor 1 homolog hUpf1
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).

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Product Information
Format	Affinity Purified
Control	Lamda phosphatase treated and untreated Jurkat RIPA lysates
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	Western Blotting Immunoprecipitation
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	Human Mouse
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	NP_002902
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	UPF1 RENT1
Modifications	Phosphorylation
Purification Method	Affinity Purfied
UniProt Number	Q92900
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.

Physicochemical Information

Dimensions

Materials Information

Toxicological Information

Safety Information according to GHS

Safety Information

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	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 2-8°C from date of receipt.

Packaging Information
Material Size	100 µg

Transport Information

Supplemental Information

Specifications

Global Trade Item Number
Catalogue Number	GTIN
07-1016	04053252683732

Documentation

Anti-phospho-Upf1 (Ser1127) Antibody MSDS

Title
Safety Data Sheet (SDS)

Anti-phospho-Upf1 (Ser1127) Antibody Certificates of Analysis

Title	Lot Number
Anti-phospho-Upf1 (Ser1127)	2479827
Anti-phospho-Upf1 (Ser1127)	2842541
Anti-phospho-Upf1 (Ser1127) - 2391155	2391155
Anti-phospho-Upf1 (Ser1127) - NRG1868522	NRG1868522
Anti-phospho-Upf1 (Ser1127) - 2051388	2051388
Anti-phospho-Upf1 (Ser1127) - 2527574	2527574
Anti-phospho-Upf1 (Ser1127) - 3169495	3169495
Anti-phospho-Upf1 (Ser1127) - 3184113	3184113
Anti-phospho-Upf1 (Ser1127) - 3218770	3218770
Anti-phospho-Upf1 (Ser1127) - 3241225	3241225

References

Reference overview	Pub Med ID
Attenuation of nonsense-mediated mRNA decay facilitates the response to chemotherapeutics. Popp, MW; Maquat, LE Nature communications 6 6632 2015 Show Abstract Nonsense-mediated mRNA decay (NMD) limits the production of aberrant mRNAs containing a premature termination codon and also controls the levels of endogenous transcripts. Here we show that when human cells are treated with clinically used chemotherapeutic compounds, NMD activity declines partly as a result of the proteolytic production of a dominant-interfering form of the key NMD factor UPF1. Production of cleaved UPF1 functions to upregulate genes involved in the response to apoptotic stresses. The biological consequence is the promotion of cell death. Combined exposure of cells to a small-molecule inhibitor of NMD, NMDI-1, and the chemotherapeutic doxorubicin leads to enhanced cell death, while inhibiting UPF1 cleavage protects cells from doxorubicin challenge. We propose a model to explain why the expression levels of genes producing mRNAs of diverse structure that encode proteins of diverse function are under the purview of NMD.	25808464

Reference overview

Pub Med ID

Catalogue Number	Description
03-191	RIPAb+ Upf1 - RIP Validated Antibody and Primer Set
07-1014	Anti-Upf1 Antibody

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