Anti-p-Upf1 (Ser1127) Antibody, clone 1C4-H1 ZooMAb^® Rabbit Monoclonal

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ZooMAb^® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb^® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb^® antibodies are reliably available and ready to ship when you need them.

Clone 1C4-H1 is a ZooMAb^® rabbit recombinant monoclonal antibody that specifically detects Up-frameshift suppressor 1 homolog (Upf1) phosphorylated on serine 1127.

KLH-conjugated linear peptide corresponding to 13 amino acids surrounding phosphoserine 1127 in human Up-frameshift suppressor 1 homolog (Upf1).

Quality Control Testing

Evaluated by Western Blotting in lysate from A431 cells treated with Calyculin A and Okadaic Acid.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected phospho-Upf1(Ser1127) in lysate from overnight starved A431 cells treated with Calyculin A (50 nM) and Okadaic acid (500 nM) for 30 minutes, but not in lysate from untreated cells.

Tested Applications

Peptide Inhibition Assay: Target band detection in a lysate from NIH3T3 cells treated with Calyculin A (50 nM) and Okadaic acid (500 nM) was prevented by preblocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide.

Affinity Binding Assay A representative lot of this antibody bound p-Upf1 (Ser1127) peptide with a KD of 5.6 x 10-7 in an affinity binding assay.

Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected phospho-Upf1 (Ser1127) in A431cells serum starved overnight then treated with Calyculin A (50 nM) and Okadaic Acid (500 nM) for 30 min.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

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 an RNA-dependent helicase and ATPase that is 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). Two isoforms of UPF1 have been described that are produced by alternative splicing. This ZooMAb^® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonal (Ref.: Chiu, S-Y., et al. (2003). RNA. 9(1); 77-87; Yamashita, A., et al. (2001). Genes Dev. 15(17); 2215-2218).

Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.

300 µg/mL after reconstitution at 25 µL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.

ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany

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.