dUTPase Recombinant Rabbit Monoclonal Antibody [JE66-11]
cat.: HA721389
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IHC-P, IF-Cell, FC
Clonality: Monoclonal
Clone number: JE66-11
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 27 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human dUTPase aa 153-252 / 252.
Positive control: HeLa cell lysate, A549 cell lysate, Jurkat cell lysate, LO2 cell lysate, HepG2 cell lysate, K-562 cell lysate, HEK-293 cell lysate, A431 cell lysate, human liver tissue lysate, human esophagus tissue, human spleen tissue, HCT116.
Subcellular location: Nucleus; Mitochondrion.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  FC

1:1,000-1:2,000
1:1,000
1:100
1:500-1:1,000
Uniprot #: SwissProt: P33316 Human
Alternative names: Deoxyuridine 5' triphosphate nucleotidohydrolase, nuclear isoform Deoxyuridine 5'-triphosphate nucleotidohydrolase, mitochondrial Deoxyuridine triphosphatase dut DUT_HUMAN dUTP nucleotidohydrolase dUTP pyrophosphatase dUTPase FLJ20622
Images
HA721389_1.jpg Fig1: Western blot analysis of dUTPase on different lysates with Rabbit anti-dUTPase antibody (HA721389) at 1/1,000 dilution.

Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: A549 cell lysate (15 µg/Lane)
Lane 3: Jurkat cell lysate (15 µg/Lane)
Lane 4: LO2 cell lysate (15 µg/Lane)
Lane 5: HepG2 cell lysate (15 µg/Lane)
Lane 6: K-562 cell lysate (15 µg/Lane)
Lane 7: HEK-293 cell lysate (15 µg/Lane)
Lane 8: A431 cell lysate (15 µg/Lane)
Lane 9: Human liver tissue lysate (30 µg/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 (HA721389) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
HA721389_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human esophagus tissue with Rabbit anti-dUTPase antibody (HA721389) 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 (HA721389) 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.
HA721389_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-dUTPase antibody (HA721389) 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 (HA721389) 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.
HA721389_4.jpg Fig4: Immunocytochemistry analysis of HCT116 cells labeling dUTPase with Rabbit anti-dUTPase antibody (HA721389) 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 (HA721389) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA721389_5.jpg Fig5: Flow cytometric analysis of HCT116 cells labeling dUTPase.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721389, 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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.