dUTPase Antibody (JE66-11)

Western Blot: dUTPase Antibody (JE66-11) [NBP3-32281] - Western blot analysis of dUTPase on different lysates with Rabbit anti-dUTPase antibody (NBP3-32281) at 1/1,000 dilution. Lane 1: HeLa cell lysate (15 ug/Lane) Lane 2: A549 cell lysate (15 ug/Lane) Lane 3: Jurkat cell lysate (15 ug/Lane) Lane 4: LO2 cell lysate (15 ug/Lane) Lane 5: HepG2 cell lysate (15 ug/Lane) Lane 6: K-562 cell lysate (15 ug/Lane) Lane 7: HEK-293 cell lysate (15 ug/Lane) Lane 8: A431 cell lysate (15 ug/Lane) Lane 9: Human liver tissue lysate (30 ug/Lane) Predicted band size: 27 kDa Observed band size: 20 kDa Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32281) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.

Immunohistochemistry: dUTPase Antibody (JE66-11) [NBP3-32281] - Immunohistochemical analysis of paraffin-embedded human esophagus tissue with Rabbit anti-dUTPase antibody (NBP3-32281) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32281) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunocytochemistry/ Immunofluorescence: dUTPase Antibody (JE66-11) [NBP3-32281] - Immunocytochemistry analysis of HCT116 cells labeling dUTPase with Rabbit anti-dUTPase antibody (NBP3-32281) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-dUTPase antibody (NBP3-32281) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.

Flow Cytometry: dUTPase Antibody (JE66-11) [NBP3-32281] - Flow cytometric analysis of HCT116 cells labeling dUTPase. Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32281, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

• Reactivity: Hu
• Applications: WB, Flow, ICC/IF, IHC
• Clone: JE66-11
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

dUTPase Antibody (JE66-11) Summary

• Description: Novus Biologicals Rabbit dUTPase Antibody (JE66-11) (NBP3-32281) is a recombinant monoclonal antibody validated for use in IHC, WB, Flow and ICC/IF. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: Recombinant protein within Human dUTPase aa 153-252 / 252. (Uniprot: P33316)
• Localization: Nucleus; Mitochondrion.
• Source: HEK293
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: DUT
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:500-1:1000
  • Immunocytochemistry/ Immunofluorescence 1:100
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:1000
  • Western Blot 1:1000-1:2000
• Theoretical MW: 27 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for dUTPase Antibody (JE66-11)

• deoxyuridine triphosphatase
• dUTP pyrophosphatasedUTP nucleotidohydrolase
• dUTPasedeoxyuridine 5'-triphosphate nucleotidohydrolase, mitochondrial
• EC 3.6.1.23
• FLJ20622

Background

dUTPase encodes an essential enzyme of nucleotide metabolism. The encoded protein forms a ubiquitous, homotetrameric enzyme that hydrolyzes dUTP to dUMP and pyrophosphate. This reaction serves two cellular purposes: providing a precursor (dUMP) for the synthesis of thymine nucleotides needed for DNA replication, and limiting intracellular pools of dUTP. Elevated levels of dUTP lead to increased incorporation of uracil into DNA, which induces extensive excision repair mediated by uracil glycosylase. This repair process, resulting in the removal and reincorporation of dUTP, is self-defeating and leads to DNA fragmentation and cell death. Alternative splicing of this gene leads to different isoforms that localize to either the mitochondrion or nucleus. A related pseudogene is located on chromosome 19.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: DUT