Nucleoside Phosphorylase Rabbit Polyclonal Antibody
cat.: HA500062
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Cell, FC
Clonality: Polyclonal
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: 1*TBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Immunogen affinity purified.
Molecular weight: Predicted band size: 32 kDa
Isotype: IgG
Immunogen: Recombinant protein within human Nucleoside Phosphorylase aa 1-200.
Positive control: K562 cell lysate, human placenta tissue, human small intestine tissue, Jurkat, NIH/3T3, PC-12.
Subcellular location: Cytoplasm.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  FC
1:20,000
1:200-1:1,000
1:250
1:1,000
Uniprot #: SwissProt: P00491 Human | P23492 Mouse | P85973 Rat
Alternative names: FLJ94043 FLJ97288 FLJ97312 Inosine phosphorylase Inosine-guanosine phosphorylase MGC117396 MGC125915 MGC125916 NP Np1 Nucleoside phosphorylase PNP Pnp1 PNPH_HUMAN PRO1837 PUNP Purine nucleoside orthophosphate ribosyltransferase Purine nucleoside phosphorylase 5a Purine nucleoside phosphorylase
Images
HA500062_1.jpg Fig1: Western blot analysis of Nucleoside Phosphorylase on K-562 cell lysate with Rabbit anti-Nucleoside Phosphorylase antibody (HA500062) at 1/20,000 dilution.

Lysates/proteins at 20 µg/Lane.
Exposure time: 60 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA500062, 1/20,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 32 kDa
Observed band size: 32 kDa
HA500062_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-Nucleoside Phosphorylase antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA500062, 1/800) for 30 minutes 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.
HA500062_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human small intestine tissue using anti-Nucleoside Phosphorylase antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA500062, 1/800) for 30 minutes 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.
HA500062_4.jpg Fig4: Immunocytochemistry analysis of Jurkat cells labeling Nucleoside Phosphorylase with Rabbit anti-Nucleoside Phosphorylase antibody (HA500062) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Nucleoside Phosphorylase antibody (HA500062) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA500062_5.jpg Fig5: Immunocytochemistry analysis of NIH/3T3 cells labeling Nucleoside Phosphorylase with Rabbit anti-Nucleoside Phosphorylase antibody (HA500062) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Nucleoside Phosphorylase antibody (HA500062) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA500062_6.jpg Fig6: Immunocytochemistry analysis of PC-12 cells labeling Nucleoside Phosphorylase with Rabbit anti-Nucleoside Phosphorylase antibody (HA500062) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Nucleoside Phosphorylase antibody (HA500062) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA500062_7.jpg Fig7: Flow cytometric analysis of Jurkat cells labeling Nucleoside Phosphorylase.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA500062, 1/1,000) (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).
HA500062_8.jpg Fig8: Flow cytometric analysis of NIH/3T3 cells labeling Nucleoside Phosphorylase.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA500062, 1/1,000) (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).
HA500062_9.jpg Fig9: Flow cytometric analysis of PC-12 cells labeling Nucleoside Phosphorylase.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA500062, 1/1,000) (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).
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