![Western Blot: p19ARF/CDKN2A Antibody (5-C3-1) [NB200-174] - Whole cell protein from mouse Neuro2A and MEF cells was separated on a 4-20% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/mL anti-p19ARF in block buffer and detected with an anti-mouse HRP secondary antibody using chemiluminescence.](http://images.novusbio.com/fullsize/p19ARF-CDKN2A-Antibody-5-C3-1-Western-Blot-NB200-174-img0011.jpg "Western Blot: p19ARF/CDKN2A Antibody (5-C3-1) [NB200-174] - Whole cell protein from mouse Neuro2A and MEF cells was separated on a 4-20% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/mL anti-p19ARF in block buffer and detected with an anti-mouse HRP secondary antibody using chemiluminescence.")
| Reactivity | Hu, Mu, SyHa(-)Species Glossary |
| Applications | WB, Flow, ICC/IF, IHC, IP, CyTOF-ready, KO |
| Clone | 5-C3-1 |
| Clonality | Monoclonal |
| Host | Rat |
| Conjugate | Unconjugated |
| Format | BSA Free |
| Concentration | 1.0 mg/ml |
| Immunogen | A synthetic peptide containing amino acids 54-75 of the murine p19ARF/CDKN2A. [UniProt# Q64364] |
| Epitope | The epitope recognized is between amino acids 62-75 of murine p19ARF/CDKN2A. |
| Isotype | IgG2b |
| Clonality | Monoclonal |
| Host | Rat |
| Gene | Cdkn2a |
| Purity | Protein A 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 | In Western blot a band can be seen at ~19 kDa. 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. |
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| Theoretical MW | 19 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. |
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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 |
| Preservative | 0.05% Sodium Azide |
| Concentration | 1.0 mg/ml |
| Purity | Protein A purified |
Secondary Antibodies |
Isotype Controls |
Research Areas for p19ARF/CDKN2A Antibody (NB200-174)Find related products by research area.
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| Gene Symbol | Cdkn2a |
![Immunohistochemistry: p19ARF/CDKN2A Antibody (5-C3-1) [NB200-174] - Staining of sections from a wild type mouse testis taken at P11 were probed with NB200-174 (green). Sections were stained with DAPI to reveal the position of cell nuclei (blue).](http://images.novusbio.com/fullsize/p19ARF-CDKN2A-Antibody-5-C3-1-Immunohistochemistry-NB200-174-img0008.jpg "Immunohistochemistry: p19ARF/CDKN2A Antibody (5-C3-1) [NB200-174] - Staining of sections from a wild type mouse testis taken at P11 were probed with NB200-174 (green). Sections were stained with DAPI to reveal the position of cell nuclei (blue).")
![Flow (Intracellular): p19ARF/CDKN2A Antibody (5-C3-1) [NB200-174] - An intracellular stain was performed on HEK293 cells with p14ARF (5-C3-1) antibody NB200-174 (blue) and a matched isotype control MAB0061 (orange) [A]. An intracellular stain was also performed on THP-1 (known homozygous deletion of gene, PMID: 19635138) cells to ensure antibody specificity [B]. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Goat Anti-Rat F(ab)2 IgG (H+L) APC-conjugated Antibody (F0113, R&D Systems). *Note: This antibody recognizes both human (p14ARF) and mouse (p19ARF) versions of the protein.](http://images.novusbio.com/fullsize/p19ARF-CDKN2A-Antibody-5-C3-1-Flow-Intracellular-NB200-174-img0010.jpg "Flow (Intracellular): p19ARF/CDKN2A Antibody (5-C3-1) [NB200-174] - An intracellular stain was performed on HEK293 cells with p14ARF (5-C3-1) antibody NB200-174 (blue) and a matched isotype control MAB0061 (orange) [A]. An intracellular stain was also performed on THP-1 (known homozygous deletion of gene, PMID: 19635138) cells to ensure antibody specificity [B]. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Goat Anti-Rat F(ab)2 IgG (H+L) APC-conjugated Antibody (F0113, R&D Systems). *Note: This antibody recognizes both human (p14ARF) and mouse (p19ARF) versions of the protein.")
![Western Blot: p19ARF/CDKN2A Antibody (5-C3-1) [NB200-174] - Aberrantly upregulated p19ARF/CDKN2A antagonizes TFH differentiation and survival. p19ARF/CDKN2A and Bcl6 interaction capacity. WT or mutant p19ARF/CDKN2A was co-transfected with WT Bcl6 expression vector into 293T cells. Cell lysates were immunoprecipitated with anti-Bcl6 and then immunoblotted with anti-p19ARF/CDKN2A. Data are representative of two experiments. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-018-07853-z) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/p19ARF-CDKN2A-Antibody-5-C3-1-Western-Blot-NB200-174-img0012.jpg "Western Blot: p19ARF/CDKN2A Antibody (5-C3-1) [NB200-174] - Aberrantly upregulated p19ARF/CDKN2A antagonizes TFH differentiation and survival. p19ARF/CDKN2A and Bcl6 interaction capacity. WT or mutant p19ARF/CDKN2A was co-transfected with WT Bcl6 expression vector into 293T cells. Cell lysates were immunoprecipitated with anti-Bcl6 and then immunoblotted with anti-p19ARF/CDKN2A. Data are representative of two experiments. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-018-07853-z) licensed under a CC-BY license.")
![Western Blot: p19ARF/CDKN2A Antibody (5-C3-1) [NB200-174] - Analysis of endogenous p19Arf. Total cell lysates (25 ug) from NIH3T3 cells, which have deleted the Arf gene (Lanes 1) and from wild type mouse embryo fibroblasts (MEFs) at passage 6, which express p19Arf (Lanes 2), were resolved by SDS-PAGE.](http://images.novusbio.com/fullsize/p19ARF-CDKN2A-Antibody-5-C3-1-Western-Blot-NB200-174-img0005.jpg "Western Blot: p19ARF/CDKN2A Antibody (5-C3-1) [NB200-174] - Analysis of endogenous p19Arf. Total cell lysates (25 ug) from NIH3T3 cells, which have deleted the Arf gene (Lanes 1) and from wild type mouse embryo fibroblasts (MEFs) at passage 6, which express p19Arf (Lanes 2), were resolved by SDS-PAGE.")
![Knockout Validated: p19ARF/CDKN2A Antibody (5-C3-1) [NB200-174] - Detection of p19Arf. Wild type mouse embryo fibroblasts (MEF) at passage 5 (panels 1, 2) and NIH3T3 cells, which have deleted the Arf gene (panels 3, 4) were probed with NB200-17 and nuclear stain (overlay in panels 2 and 4).](http://images.novusbio.com/fullsize/p19ARF-CDKN2A-Antibody-5-C3-1-Knockout-Validated-NB200-174-img0009.jpg "Knockout Validated: p19ARF/CDKN2A Antibody (5-C3-1) [NB200-174] - Detection of p19Arf. Wild type mouse embryo fibroblasts (MEF) at passage 5 (panels 1, 2) and NIH3T3 cells, which have deleted the Arf gene (panels 3, 4) were probed with NB200-17 and nuclear stain (overlay in panels 2 and 4).")
![Western Blot: p19ARF/CDKN2A Antibody (5-C3-1) - BSA Free [NB200-174] - Phosphorylation of NPM-Ser48 regulates the localization of NPM & p19A(A) Npm−/−, p53−/−double null MEF were infected with pBabe retrovirus expressing FLAG-tagged-myr-AKT1 in combination with NPM-WT or NPM-S48A as indicated. Cells were fixed & stained with DAPI & anti-NPM (left) or anti-phospho-S48-NPM (pS48-NPM). (B) NPM immunoprecipitates & whole cell lysates from Npm−/−;p53−/− MEFs expressing human NPM or NPM-S48A were probed with the indicated antibodies.(C) Graph, quantification of p19ARF staining intensity in confocal images using ImageJ. (D) Npm−/−, p53−/−double null MEF were infected with pBABE retrovirus expressing FLAG-tagged-myr-AKT1 in combination with NPM-WT, NPM-S48A or S48E as indicated. Cells were fixed & stained with DAPI, anti-NPM & anti p19ARF. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25071014), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb200-174_rat-monoclonal-p19arf-cdkn2a-antibody-5-c3-1-31020241621829.jpg "Western Blot: p19ARF/CDKN2A Antibody (5-C3-1) - BSA Free [NB200-174] - Phosphorylation of NPM-Ser48 regulates the localization of NPM & p19A(A) Npm−/−, p53−/−double null MEF were infected with pBabe retrovirus expressing FLAG-tagged-myr-AKT1 in combination with NPM-WT or NPM-S48A as indicated. Cells were fixed & stained with DAPI & anti-NPM (left) or anti-phospho-S48-NPM (pS48-NPM). (B) NPM immunoprecipitates & whole cell lysates from Npm−/−;p53−/− MEFs expressing human NPM or NPM-S48A were probed with the indicated antibodies.(C) Graph, quantification of p19ARF staining intensity in confocal images using ImageJ. (D) Npm−/−, p53−/−double null MEF were infected with pBABE retrovirus expressing FLAG-tagged-myr-AKT1 in combination with NPM-WT, NPM-S48A or S48E as indicated. Cells were fixed & stained with DAPI, anti-NPM & anti p19ARF. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25071014), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Western Blot: p19ARF/CDKN2A Antibody (5-C3-1) - BSA Free [NB200-174] - Inhibition of AKT promotes enhanced MDM2 activity via the increased association between NPM & p14ARF(A) Npm−/−, p53−/−double null MEF were infected with pBABE retrovirus empty vector & pBABE expressing FLAG-tagged-NPM-WT, NPM-S48A or S48E as indicated. Immunopurification of NPM was done by pulling down with the Flag tag (middle panel) followed by elution of complexes by the Flag peptide & subsequent immunopurification of endogenous MDM2 (lower panel). (B) Nuclear immunoprecipitates of MDM2 from T24 cells treated with MK-2206 (5 μM, 24 hrs). Immunoprecipitates & lysates were blotted with the indicated antibodies. (C) T24 cells were treated with MK-2206 (5 μM) as indicated. p14ARF was immunoprecipitated from whole cell lysates & nuclear extracts & the association with NPM & MDM2 determined by western blot. Immunoprecipitates & lysates were blotted with the indicated antibodies. (D) MDM2 & (E) p53 ubiquinitation assay in H1299 cells transfected with wild type p53, HA-tagged ubiquitin & treated for 16 hrs with DMSO, MK-2206 (5 μM) or Nutlin3A (5 μM) as indicated. Immunoprecipitates & whole cell lysates were probed with the indicated antibodies. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25071014), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb200-174_rat-monoclonal-p19arf-cdkn2a-antibody-5-c3-1-310202416205122.jpg "Western Blot: p19ARF/CDKN2A Antibody (5-C3-1) - BSA Free [NB200-174] - Inhibition of AKT promotes enhanced MDM2 activity via the increased association between NPM & p14ARF(A) Npm−/−, p53−/−double null MEF were infected with pBABE retrovirus empty vector & pBABE expressing FLAG-tagged-NPM-WT, NPM-S48A or S48E as indicated. Immunopurification of NPM was done by pulling down with the Flag tag (middle panel) followed by elution of complexes by the Flag peptide & subsequent immunopurification of endogenous MDM2 (lower panel). (B) Nuclear immunoprecipitates of MDM2 from T24 cells treated with MK-2206 (5 μM, 24 hrs). Immunoprecipitates & lysates were blotted with the indicated antibodies. (C) T24 cells were treated with MK-2206 (5 μM) as indicated. p14ARF was immunoprecipitated from whole cell lysates & nuclear extracts & the association with NPM & MDM2 determined by western blot. Immunoprecipitates & lysates were blotted with the indicated antibodies. (D) MDM2 & (E) p53 ubiquinitation assay in H1299 cells transfected with wild type p53, HA-tagged ubiquitin & treated for 16 hrs with DMSO, MK-2206 (5 μM) or Nutlin3A (5 μM) as indicated. Immunoprecipitates & whole cell lysates were probed with the indicated antibodies. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25071014), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Western Blot: p19ARF/CDKN2A Antibody (5-C3-1) - BSA Free [NB200-174] - Loss of P19Arf enhances tumor growth & primary lung fibroblasts induce p19Arf in response to leucine deprivation (LD). (A) Graph of sarcoma (SKPY) tumor volume on the indicated days. SKPY cells were injected into the flank of WT & p19Arf-null mice. Tumor volume was measured by caliper. N=7 mice/WT cohort & N=15 mice/p19Arf−/− cohort. (B) Western blot for p19Arf protein expression in WT & Arf-null fibroblasts after LD for the indicated days. Densitometric quantification of p19Arf protein expression relative to beta -tubulin is shown on right. (C) qPCR for p19Arf mRNA in WT or p19Arf−/− fibroblasts during LD for the indicated days. (D) Representative immunofluorescence images for P19Arf subcellular localization on the indicated days after LD in WT or p19Arf−/− ALFs. Quantification of nuclear p19Arf is shown in graph on right. (E) Western blot analysis of phospho-eIF2 alpha (P-eIF2 alpha ) & total eIF2 alpha expression in WT & p19Arf-null murine ALFs during LD for the indicated days. Graph quantifies intensity of P-eIF2 alpha expression relative to day 0. beta -Tubulin used as loading control. (F) Western blot analysis of P-eIF2 alpha expression in WT & p19Arf−/− ALFs upon treatment with a GCN2 inhibitor (GCN2-IN-1; A-92 1 µM in DMSO) during overnight LD. Actin used as a loading control. Quantification of P-eIF2 alpha expression relative to total eIF2 alpha . N=3; *, P<0.05; ns, not significant. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/34994382), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb200-174_rat-monoclonal-p19arf-cdkn2a-antibody-5-c3-1-310202416205128.jpg "Western Blot: p19ARF/CDKN2A Antibody (5-C3-1) - BSA Free [NB200-174] - Loss of P19Arf enhances tumor growth & primary lung fibroblasts induce p19Arf in response to leucine deprivation (LD). (A) Graph of sarcoma (SKPY) tumor volume on the indicated days. SKPY cells were injected into the flank of WT & p19Arf-null mice. Tumor volume was measured by caliper. N=7 mice/WT cohort & N=15 mice/p19Arf−/− cohort. (B) Western blot for p19Arf protein expression in WT & Arf-null fibroblasts after LD for the indicated days. Densitometric quantification of p19Arf protein expression relative to beta -tubulin is shown on right. (C) qPCR for p19Arf mRNA in WT or p19Arf−/− fibroblasts during LD for the indicated days. (D) Representative immunofluorescence images for P19Arf subcellular localization on the indicated days after LD in WT or p19Arf−/− ALFs. Quantification of nuclear p19Arf is shown in graph on right. (E) Western blot analysis of phospho-eIF2 alpha (P-eIF2 alpha ) & total eIF2 alpha expression in WT & p19Arf-null murine ALFs during LD for the indicated days. Graph quantifies intensity of P-eIF2 alpha expression relative to day 0. beta -Tubulin used as loading control. (F) Western blot analysis of P-eIF2 alpha expression in WT & p19Arf−/− ALFs upon treatment with a GCN2 inhibitor (GCN2-IN-1; A-92 1 µM in DMSO) during overnight LD. Actin used as a loading control. Quantification of P-eIF2 alpha expression relative to total eIF2 alpha . N=3; *, P<0.05; ns, not significant. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/34994382), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Immunocytochemistry/ Immunofluorescence: p19ARF/CDKN2A Antibody (5-C3-1) - BSA Free [NB200-174] - Phosphorylation of NPM-Ser48 regulates the localization of NPM & p19A(A) Npm−/−, p53−/−double null MEF were infected with pBabe retrovirus expressing FLAG-tagged-myr-AKT1 in combination with NPM-WT or NPM-S48A as indicated. Cells were fixed & stained with DAPI & anti-NPM (left) or anti-phospho-S48-NPM (pS48-NPM). (B) NPM immunoprecipitates & whole cell lysates from Npm−/−;p53−/− MEFs expressing human NPM or NPM-S48A were probed with the indicated antibodies.(C) Graph, quantification of p19ARF staining intensity in confocal images using ImageJ. (D) Npm−/−, p53−/−double null MEF were infected with pBABE retrovirus expressing FLAG-tagged-myr-AKT1 in combination with NPM-WT, NPM-S48A or S48E as indicated. Cells were fixed & stained with DAPI, anti-NPM & anti p19ARF. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25071014), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb200-174_rat-monoclonal-p19arf-cdkn2a-antibody-5-c3-1-310202416203717.jpg "Immunocytochemistry/ Immunofluorescence: p19ARF/CDKN2A Antibody (5-C3-1) - BSA Free [NB200-174] - Phosphorylation of NPM-Ser48 regulates the localization of NPM & p19A(A) Npm−/−, p53−/−double null MEF were infected with pBabe retrovirus expressing FLAG-tagged-myr-AKT1 in combination with NPM-WT or NPM-S48A as indicated. Cells were fixed & stained with DAPI & anti-NPM (left) or anti-phospho-S48-NPM (pS48-NPM). (B) NPM immunoprecipitates & whole cell lysates from Npm−/−;p53−/− MEFs expressing human NPM or NPM-S48A were probed with the indicated antibodies.(C) Graph, quantification of p19ARF staining intensity in confocal images using ImageJ. (D) Npm−/−, p53−/−double null MEF were infected with pBABE retrovirus expressing FLAG-tagged-myr-AKT1 in combination with NPM-WT, NPM-S48A or S48E as indicated. Cells were fixed & stained with DAPI, anti-NPM & anti p19ARF. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25071014), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Flow Cytometry: p19ARF/CDKN2A Antibody (5-C3-1) - BSA Free - Conducted the antibody trial with two different dilutions of p19ARF/CDKN2A Antibody (5-C3-1) [PE] (Catalog # NB200-174PE) (1:500 and 1:100) and compared them to an IgG control which was also PE labelled. Image from a verified customer review.](http://images.novusbio.com/fullsize/nb200-174_rat-monoclonal-p19arf-cdkn2a-antibody-5-c3-1-41220241042101.png "Flow Cytometry: p19ARF/CDKN2A Antibody (5-C3-1) - BSA Free - Conducted the antibody trial with two different dilutions of p19ARF/CDKN2A Antibody (5-C3-1) [PE] (Catalog # NB200-174PE) (1:500 and 1:100) and compared them to an IgG control which was also PE labelled. Image from a verified customer review.")
![Flow Cytometry: p19ARF/CDKN2A Antibody (5-C3-1) - BSA Free - Conducted the antibody trial with two different dilutions of p19ARF/CDKN2A Antibody (5-C3-1) [PE] (Catalog # NB200-174PE) (1:500 and 1:100) and compared them to an IgG control which was also PE labelled. Image from a verified customer review.](http://images.novusbio.com/fullsize/nb200-174_rat-monoclonal-p19arf-cdkn2a-antibody-5-c3-1-312202414314910.png "Flow Cytometry: p19ARF/CDKN2A Antibody (5-C3-1) - BSA Free - Conducted the antibody trial with two different dilutions of p19ARF/CDKN2A Antibody (5-C3-1) [PE] (Catalog # NB200-174PE) (1:500 and 1:100) and compared them to an IgG control which was also PE labelled. Image from a verified customer review.")
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