![Immunocytochemistry/Immunofluorescence: NT5C Antibody [NBP1-84563] - Staining of human cell line U-2 OS shows localization to cytosol. Antibody staining is shown in green.](http://images.novusbio.com/fullsize/NT5C-Antibody-Immunocytochemistry-Immunofluorescence-NBP1-84563-img0011.jpg "Immunocytochemistry/Immunofluorescence: NT5C Antibody [NBP1-84563] - Staining of human cell line U-2 OS shows localization to cytosol. Antibody staining is shown in green.")
| Reactivity | HuSpecies Glossary |
| Applications | WB, Simple Western, ICC/IF, IHC, KD |
| Clonality | Polyclonal |
| Host | Rabbit |
| Conjugate | Unconjugated |
| Format | BSA Free |
| Immunogen | This antibody was developed against Recombinant Protein corresponding to amino acids: VLADFEAGLLRGFRRRFPEEPHVPLEQRRGFLAREQYRALRPDLADKVASVYEAPGFFLDLEPIP |
| Isotype | IgG |
| Clonality | Polyclonal |
| Host | Rabbit |
| Gene | NT5C |
| Purity | Immunogen affinity purified |
| Innovator's Reward | Test in a species/application not listed above to receive a full credit towards a future purchase. |
| Dilutions |
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| Application Notes | ICC/IF Fixation Permeabilization: Use PFA/Triton X-100. IHC-Paraffin HIER pH6 retrieval is recommended. |
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| Publications |
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| Storage | Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. |
| Buffer | PBS (pH 7.2) and 40% Glycerol |
| Preservative | 0.02% Sodium Azide |
| Purity | Immunogen affinity purified |
![Immunohistochemistry EGFR Antibody [Unconjugated]](https://images.novusbio.com/images2/EGF_R_AF231_Immunohistochemistry_20892.jpg)
![Immunocytochemistry EGFR Antibody [Unconjugated]](https://images.novusbio.com/images2/EGF_R_AF231_Immunocytochemistry__Immunofluorescence_21143.jpg)
![Western Blot EGFR Antibody [Unconjugated]](https://images.novusbio.com/images2/EGF_R_AF231_Western_Blot_19925.jpg)
| Publication using NBP1-84563 | Applications | Species |
|---|---|---|
| Moniz LS, Surinova S, Ghazaly E et al. Phosphoproteomic comparison of Pik3ca and Pten signalling identifies the nucleotidase NT5C as a novel AKT substrate Sci Rep 2017-01-06 [PMID: 28059163] (WB, Human) | WB | Human |
Secondary Antibodies |
Isotype Controls |
The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.
| Gene Symbol | NT5C |
![Simple Western: NT5C Antibody [NBP1-84563] - Simple Western lane view shows a specific band for NT5C in 0.2 mg/ml of RT-4 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/NT5C-Antibody-Simple-Western-NBP1-84563-img0007.jpg "Simple Western: NT5C Antibody [NBP1-84563] - Simple Western lane view shows a specific band for NT5C in 0.2 mg/ml of RT-4 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.")
![Simple Western: NT5C Antibody [NBP1-84563] - Simple Western lane view shows a specific band for NT5C in 0.2 mg/ml of U-2OS lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/NT5C-Antibody-Simple-Western-NBP1-84563-img0008.jpg "Simple Western: NT5C Antibody [NBP1-84563] - Simple Western lane view shows a specific band for NT5C in 0.2 mg/ml of U-2OS lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.")
![Simple Western: NT5C Antibody [NBP1-84563] - Electropherogram image(s) of corresponding Simple Western lane view. NT5C antibody was used at 1:20 dilution on U-2OS lysates(s).](http://images.novusbio.com/fullsize/NT5C-Antibody-Simple-Western-NBP1-84563-img0009.jpg "Simple Western: NT5C Antibody [NBP1-84563] - Electropherogram image(s) of corresponding Simple Western lane view. NT5C antibody was used at 1:20 dilution on U-2OS lysates(s).")
![Western Blot: NT5C Antibody [NBP1-84563] - Regulation of NT5C S184 phosphorylation.(a) Sensitivity of S184 phosphorylation to PI3K pathway inhibitors. Primary MEFs were treated with A66 (3 μM), MK-2206 (1 μM), Rapamycin (20 nM), U0126 (10 μM) or DMSO as control for 3 h or overnight (o/n) for 16–18 h before lysis. (left) Lysates were immunoblotted as indicated. (right) Quantification of 4 independent experiments. Values are expressed relative to DMSO treated Pik3caH1047R/WT cells. Error bars are sem, **p < 0.01. (b) Sensitivity of S184 phosphorylation to growth factor deprivation. Indicated primary MEFs were grown in 10% FBS or starved overnight in 0.1% FBS. (top) Lysates were immunoblotted as indicated. (bottom) Quantification of 5 independent experiments. Values are expressed relative to Pik3caH1047R/WT cells grown in 10% FBS. Error bars are sem, ***p < 0.001. (c) Sensitivity of S184 phosphorylation to growth factor stimulation. Primary MEFs were starved overnight in 0.1% FBS & stimulated with FBS (10%), Insulin (100 nM) or EGF (100 ng/ml) for the indicated times before lysis. DMSO or MK-2206 (1 μM) was added at same time as stimuli. (top) Lysates were immunoblotted as indicated. (bottom) Quantification of 3–5 independent experiments. Values are expressed relative to time 0 for each stimuli. Error bars are sem. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28059163), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-84563_rabbit-polyclonal-nt5c-antibody-31020241612237.jpg "Western Blot: NT5C Antibody [NBP1-84563] - Regulation of NT5C S184 phosphorylation.(a) Sensitivity of S184 phosphorylation to PI3K pathway inhibitors. Primary MEFs were treated with A66 (3 μM), MK-2206 (1 μM), Rapamycin (20 nM), U0126 (10 μM) or DMSO as control for 3 h or overnight (o/n) for 16–18 h before lysis. (left) Lysates were immunoblotted as indicated. (right) Quantification of 4 independent experiments. Values are expressed relative to DMSO treated Pik3caH1047R/WT cells. Error bars are sem, **p < 0.01. (b) Sensitivity of S184 phosphorylation to growth factor deprivation. Indicated primary MEFs were grown in 10% FBS or starved overnight in 0.1% FBS. (top) Lysates were immunoblotted as indicated. (bottom) Quantification of 5 independent experiments. Values are expressed relative to Pik3caH1047R/WT cells grown in 10% FBS. Error bars are sem, ***p < 0.001. (c) Sensitivity of S184 phosphorylation to growth factor stimulation. Primary MEFs were starved overnight in 0.1% FBS & stimulated with FBS (10%), Insulin (100 nM) or EGF (100 ng/ml) for the indicated times before lysis. DMSO or MK-2206 (1 μM) was added at same time as stimuli. (top) Lysates were immunoblotted as indicated. (bottom) Quantification of 3–5 independent experiments. Values are expressed relative to time 0 for each stimuli. Error bars are sem. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28059163), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Western Blot: NT5C Antibody [NBP1-84563] - Impact of S184 phosphorylation of NT5C catalytic activity.(a) S184 phosphorylation does not regulate NT5C nucleotidase activity in vitro. (left) Immunoprecipitates of Flag-NT5C ectopically expressed in HEK293 cells were incubated with 5 mM of the indicated nucleotides. Phosphate release was measured using a malachite green colorimetric assay & expressed as a percent of total nucleotide. The experiment was performed in duplicate & repeated 3 times independently. Error bars are sem. (right) Representative immunoblot from experimental cells. (b) Cells expressing Pik3caH1047R have elevated dNTP levels. (left, middle) Nucleotides were extracted from primary MEFs & analysed by UPLC-MS/MS. The experiment was performed in triplicate & repeated 4 times independently. Error bars are sem, *p < 0.05, **p < 0.01. (right) Representative immunoblot from experimental cells. (c) Effect of NT5C knockdown on cellular dCTP levels. (left) Nucleotides were extracted from primary MEFs, stably expressing indicated siRNA, & analysed by UPLC-MS/MS. The experiment was performed in triplicate & repeated 3 times independently. Error bars are sem. (right) Representative immunoblot from experimental cells. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28059163), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-84563_rabbit-polyclonal-nt5c-antibody-310202416121989.jpg "Western Blot: NT5C Antibody [NBP1-84563] - Impact of S184 phosphorylation of NT5C catalytic activity.(a) S184 phosphorylation does not regulate NT5C nucleotidase activity in vitro. (left) Immunoprecipitates of Flag-NT5C ectopically expressed in HEK293 cells were incubated with 5 mM of the indicated nucleotides. Phosphate release was measured using a malachite green colorimetric assay & expressed as a percent of total nucleotide. The experiment was performed in duplicate & repeated 3 times independently. Error bars are sem. (right) Representative immunoblot from experimental cells. (b) Cells expressing Pik3caH1047R have elevated dNTP levels. (left, middle) Nucleotides were extracted from primary MEFs & analysed by UPLC-MS/MS. The experiment was performed in triplicate & repeated 4 times independently. Error bars are sem, *p < 0.05, **p < 0.01. (right) Representative immunoblot from experimental cells. (c) Effect of NT5C knockdown on cellular dCTP levels. (left) Nucleotides were extracted from primary MEFs, stably expressing indicated siRNA, & analysed by UPLC-MS/MS. The experiment was performed in triplicate & repeated 3 times independently. Error bars are sem. (right) Representative immunoblot from experimental cells. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28059163), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Western Blot: NT5C Antibody [NBP1-84563] - Regulation of NT5C S184 phosphorylation.(a) Sensitivity of S184 phosphorylation to PI3K pathway inhibitors. Primary MEFs were treated with A66 (3 μM), MK-2206 (1 μM), Rapamycin (20 nM), U0126 (10 μM) or DMSO as control for 3 h or overnight (o/n) for 16–18 h before lysis. (left) Lysates were immunoblotted as indicated. (right) Quantification of 4 independent experiments. Values are expressed relative to DMSO treated Pik3caH1047R/WT cells. Error bars are sem, **p < 0.01. (b) Sensitivity of S184 phosphorylation to growth factor deprivation. Indicated primary MEFs were grown in 10% FBS or starved overnight in 0.1% FBS. (top) Lysates were immunoblotted as indicated. (bottom) Quantification of 5 independent experiments. Values are expressed relative to Pik3caH1047R/WT cells grown in 10% FBS. Error bars are sem, ***p < 0.001. (c) Sensitivity of S184 phosphorylation to growth factor stimulation. Primary MEFs were starved overnight in 0.1% FBS & stimulated with FBS (10%), Insulin (100 nM) or EGF (100 ng/ml) for the indicated times before lysis. DMSO or MK-2206 (1 μM) was added at same time as stimuli. (top) Lysates were immunoblotted as indicated. (bottom) Quantification of 3–5 independent experiments. Values are expressed relative to time 0 for each stimuli. Error bars are sem. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28059163), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-84563_rabbit-polyclonal-nt5c-antibody-310202416113523.jpg "Western Blot: NT5C Antibody [NBP1-84563] - Regulation of NT5C S184 phosphorylation.(a) Sensitivity of S184 phosphorylation to PI3K pathway inhibitors. Primary MEFs were treated with A66 (3 μM), MK-2206 (1 μM), Rapamycin (20 nM), U0126 (10 μM) or DMSO as control for 3 h or overnight (o/n) for 16–18 h before lysis. (left) Lysates were immunoblotted as indicated. (right) Quantification of 4 independent experiments. Values are expressed relative to DMSO treated Pik3caH1047R/WT cells. Error bars are sem, **p < 0.01. (b) Sensitivity of S184 phosphorylation to growth factor deprivation. Indicated primary MEFs were grown in 10% FBS or starved overnight in 0.1% FBS. (top) Lysates were immunoblotted as indicated. (bottom) Quantification of 5 independent experiments. Values are expressed relative to Pik3caH1047R/WT cells grown in 10% FBS. Error bars are sem, ***p < 0.001. (c) Sensitivity of S184 phosphorylation to growth factor stimulation. Primary MEFs were starved overnight in 0.1% FBS & stimulated with FBS (10%), Insulin (100 nM) or EGF (100 ng/ml) for the indicated times before lysis. DMSO or MK-2206 (1 μM) was added at same time as stimuli. (top) Lysates were immunoblotted as indicated. (bottom) Quantification of 3–5 independent experiments. Values are expressed relative to time 0 for each stimuli. Error bars are sem. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28059163), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Western Blot: NT5C Antibody [NBP1-84563] - Impact of S184 phosphorylation of NT5C catalytic activity.(a) S184 phosphorylation does not regulate NT5C nucleotidase activity in vitro. (left) Immunoprecipitates of Flag-NT5C ectopically expressed in HEK293 cells were incubated with 5 mM of the indicated nucleotides. Phosphate release was measured using a malachite green colorimetric assay & expressed as a percent of total nucleotide. The experiment was performed in duplicate & repeated 3 times independently. Error bars are sem. (right) Representative immunoblot from experimental cells. (b) Cells expressing Pik3caH1047R have elevated dNTP levels. (left, middle) Nucleotides were extracted from primary MEFs & analysed by UPLC-MS/MS. The experiment was performed in triplicate & repeated 4 times independently. Error bars are sem, *p < 0.05, **p < 0.01. (right) Representative immunoblot from experimental cells. (c) Effect of NT5C knockdown on cellular dCTP levels. (left) Nucleotides were extracted from primary MEFs, stably expressing indicated siRNA, & analysed by UPLC-MS/MS. The experiment was performed in triplicate & repeated 3 times independently. Error bars are sem. (right) Representative immunoblot from experimental cells. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28059163), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-84563_rabbit-polyclonal-nt5c-antibody-31020241612211.jpg "Western Blot: NT5C Antibody [NBP1-84563] - Impact of S184 phosphorylation of NT5C catalytic activity.(a) S184 phosphorylation does not regulate NT5C nucleotidase activity in vitro. (left) Immunoprecipitates of Flag-NT5C ectopically expressed in HEK293 cells were incubated with 5 mM of the indicated nucleotides. Phosphate release was measured using a malachite green colorimetric assay & expressed as a percent of total nucleotide. The experiment was performed in duplicate & repeated 3 times independently. Error bars are sem. (right) Representative immunoblot from experimental cells. (b) Cells expressing Pik3caH1047R have elevated dNTP levels. (left, middle) Nucleotides were extracted from primary MEFs & analysed by UPLC-MS/MS. The experiment was performed in triplicate & repeated 4 times independently. Error bars are sem, *p < 0.05, **p < 0.01. (right) Representative immunoblot from experimental cells. (c) Effect of NT5C knockdown on cellular dCTP levels. (left) Nucleotides were extracted from primary MEFs, stably expressing indicated siRNA, & analysed by UPLC-MS/MS. The experiment was performed in triplicate & repeated 3 times independently. Error bars are sem. (right) Representative immunoblot from experimental cells. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28059163), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Western Blot: NT5C Antibody [NBP1-84563] - Regulation of NT5C S184 phosphorylation.(a) Sensitivity of S184 phosphorylation to PI3K pathway inhibitors. Primary MEFs were treated with A66 (3 μM), MK-2206 (1 μM), Rapamycin (20 nM), U0126 (10 μM) or DMSO as control for 3 h or overnight (o/n) for 16–18 h before lysis. (left) Lysates were immunoblotted as indicated. (right) Quantification of 4 independent experiments. Values are expressed relative to DMSO treated Pik3caH1047R/WT cells. Error bars are sem, **p < 0.01. (b) Sensitivity of S184 phosphorylation to growth factor deprivation. Indicated primary MEFs were grown in 10% FBS or starved overnight in 0.1% FBS. (top) Lysates were immunoblotted as indicated. (bottom) Quantification of 5 independent experiments. Values are expressed relative to Pik3caH1047R/WT cells grown in 10% FBS. Error bars are sem, ***p < 0.001. (c) Sensitivity of S184 phosphorylation to growth factor stimulation. Primary MEFs were starved overnight in 0.1% FBS & stimulated with FBS (10%), Insulin (100 nM) or EGF (100 ng/ml) for the indicated times before lysis. DMSO or MK-2206 (1 μM) was added at same time as stimuli. (top) Lysates were immunoblotted as indicated. (bottom) Quantification of 3–5 independent experiments. Values are expressed relative to time 0 for each stimuli. Error bars are sem. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28059163), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-84563_rabbit-polyclonal-nt5c-antibody-310202416114732.jpg "Western Blot: NT5C Antibody [NBP1-84563] - Regulation of NT5C S184 phosphorylation.(a) Sensitivity of S184 phosphorylation to PI3K pathway inhibitors. Primary MEFs were treated with A66 (3 μM), MK-2206 (1 μM), Rapamycin (20 nM), U0126 (10 μM) or DMSO as control for 3 h or overnight (o/n) for 16–18 h before lysis. (left) Lysates were immunoblotted as indicated. (right) Quantification of 4 independent experiments. Values are expressed relative to DMSO treated Pik3caH1047R/WT cells. Error bars are sem, **p < 0.01. (b) Sensitivity of S184 phosphorylation to growth factor deprivation. Indicated primary MEFs were grown in 10% FBS or starved overnight in 0.1% FBS. (top) Lysates were immunoblotted as indicated. (bottom) Quantification of 5 independent experiments. Values are expressed relative to Pik3caH1047R/WT cells grown in 10% FBS. Error bars are sem, ***p < 0.001. (c) Sensitivity of S184 phosphorylation to growth factor stimulation. Primary MEFs were starved overnight in 0.1% FBS & stimulated with FBS (10%), Insulin (100 nM) or EGF (100 ng/ml) for the indicated times before lysis. DMSO or MK-2206 (1 μM) was added at same time as stimuli. (top) Lysates were immunoblotted as indicated. (bottom) Quantification of 3–5 independent experiments. Values are expressed relative to time 0 for each stimuli. Error bars are sem. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28059163), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Immunohistochemistry-Paraffin: NT5C Antibody [NBP1-84563] -Staining of human kidney shows moderate cytoplasmic positivity in cells in tubules.](http://images.novusbio.com/fullsize/antibody/nbp1-84563_rabbit-polyclonal-nt5c-antibody-immunohistochemistry-paraffin-1312202495154..png "Immunohistochemistry-Paraffin: NT5C Antibody [NBP1-84563] -Staining of human kidney shows moderate cytoplasmic positivity in cells in tubules.")
![Immunohistochemistry-Paraffin: NT5C Antibody [NBP1-84563] -Staining of human cerebral cortex shows moderate cytoplasmic positivity in neuronal cells.](http://images.novusbio.com/fullsize/antibody/nbp1-84563_rabbit-polyclonal-nt5c-antibody-immunohistochemistry-paraffin-1312202495245..png "Immunohistochemistry-Paraffin: NT5C Antibody [NBP1-84563] -Staining of human cerebral cortex shows moderate cytoplasmic positivity in neuronal cells.")
![Immunohistochemistry-Paraffin: NT5C Antibody [NBP1-84563] -Staining of human liver shows weak cytoplasmic positivity in hepatocytes as expected.](http://images.novusbio.com/fullsize/antibody/nbp1-84563_rabbit-polyclonal-nt5c-antibody-immunohistochemistry-paraffin-1312202495335..png "Immunohistochemistry-Paraffin: NT5C Antibody [NBP1-84563] -Staining of human liver shows weak cytoplasmic positivity in hepatocytes as expected.")
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![Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP] [NB7160] - Western blot showing vemurafenib treatment in BRAFV600E CRC cells inhibits fission mediator DRP1 with no significant effect on fusion proteins (Mfn1 & 2) using MFN-1 antibody (NBP1-51841) and corresponding secondary antibody, goat anti-rabbit IgG-HRP (NB7160). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33738242).](https://images.novusbio.com/images/Goat-anti-Rabbit-IgG-H+L-Secondary-Antibody-HRP-Western-Blot-NB7160-img0001.jpg "Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP] [NB7160] - Western blot showing vemurafenib treatment in BRAFV600E CRC cells inhibits fission mediator DRP1 with no significant effect on fusion proteins (Mfn1 & 2) using MFN-1 antibody (NBP1-51841) and corresponding secondary antibody, goat anti-rabbit IgG-HRP (NB7160). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33738242).")
![Flow Cytometry: Rabbit IgG Isotype Control [NBP2-24891] - Intracellular FACS analysis of mouse TLR6 polyclonal antibody (red), rabbit isotype control (green), RAW cells alone (shaded). Two micrograms of antibodies were used. Goat anti-rabbit FITC (Novus, 20302) was used as secondary.](https://images.novusbio.com/images/Rabbit--Mouse-IgG-Isotype-Control-Flow-Cytometry-NBP2-24891-img0001.jpg "Flow Cytometry: Rabbit IgG Isotype Control [NBP2-24891] - Intracellular FACS analysis of mouse TLR6 polyclonal antibody (red), rabbit isotype control (green), RAW cells alone (shaded). Two micrograms of antibodies were used. Goat anti-rabbit FITC (Novus, 20302) was used as secondary.")