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Primary Antibodies  >  Cell Cycle Protein Antibodies
 

p19Arf Antibody

Rat Monoclonal 5.C3.1 IgG2b
Code

200-501-891
Size

100 µg
Price

$309.00
Availability

Ships next business day.
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E-mail: orders@rockland-inc.com
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Immunofluorescence microscopy using Rockland's Protein G Purified Rat Monoclonal Anti-p19Arf antibody to detect endogenous mouse p19Arf protein present in sections of testis from a wild type mouse at stage P11, probed with 5-C3-1 MAb (green).  The section was also incubated with DAPI, to show positions of nuclei (blue). A similar section from the testis of an Arf -/- mutant mouse does not stain with this MAb (data not shown).  Tissue was perfused with 4% paraformaldehyde in PBS.  Slides were placed in a jar containing citrate buffer at pH 6.0 at 96° C for 30 min followed by blocking in 10% normal goat serum in 0.1% Triton X-100-PBS for 30 min.  Slides were incubated with 5 ug/ml antibody at 4° C overnight followed by additional of fluorochrome conjugated secondary antibody.  See Bertwistle et al. 2004 for additional details.
Western blot analysis is shown using Rockland's Protein G Purified Rat Monoclonal Anti-p19Arf antibody to detect endogenous mouse p19Arf protein present in wtMEFp6 cells (wild type mouse embryo fibroblasts passage 6).   No reactivity is observed in lysates from NIH3T3 cells, which lack the Arf gene.  Approximately 50 ?g of cell lysate was loaded on a SDS-PAGE gel, blotted onto a PVDF membrane and probed with 1.0 ?g/ml of rat-anti-p19Arf antibody (both clones 5-C3-1 and 12-A1-1 are shown).  The blot was visualized using HRP conjugated Gt-anti-Rat IgG followed by ECL reagent.  See Bertwistle et al. 2004 for details.
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Synonyms: Cyclin-dependent kinase inhibitor 2A, isoform 3 p19ARF

p19Arf Antibody Properties

Description
Anti-p19Arf (exon 2) (RAT) Monoclonal Antibody - 200-501-891
Host
Rat
Target Species
Mouse
Known Cross Reactivity
mouse, human, hamster
Clonality
Monoclonal 5.C3.1 IgG2b
Application
ELISA : 1:5,000 - 1:20,000
IF Microscopy : 1:500 - 1:2,000
Western Blot : 1:500 - 1:2,000
ImmunoPrecipitation: 1:100
Other Dilution: User Optimized
Physical State
Liquid (sterile filtered)
Shipping condition
Dry Ice
Format
IgG2b
Label
Concentration
1.0 mg/mL by UV absorbance at 280 nm
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
0.01% (w/v) Sodium Azide
Stabilizer
None
GeneName
CDKN2A
Download CofA
Lot#  

p19Arf Antibody Description

Background
The Ink4a-Arf locus encodes two tumor suppressor proteins, p16Ink4a and p19Arf.  Expression of Ink4a and Arf is regulated by distinct promoters upstream of alternative first exons whose products are spliced to a common second exon translated in alternative reading frames (from which Arf gets its name). The two genes are induced by different stress signals and can be separately mutated or silenced in tumor cells.  Targeted disruption of either or both genes in mouse leads to spontaneous tumor formation, which is accelerated by exposure of the mutant animals to chemical carcinogens or ionizing radiation.  Inactivation of ARF by mutation, deletion, or epigenetic silencing is observed in many human cancers, underscoring the role of this protein as a potent and ubiquitous tumor suppressor.  p19Arf  is activated by Myc or mutant Ras.  Once activated, p19Arf binds to the p53 negative regulator Mdm2, leading to p53 stabilization and unleashing a p53-dependent transcriptional program that triggers either cell cycle arrest or apoptosis.  p19Arf also has p53-independent inhibitory effects on cell proliferation.  p19Arf is a nucleolar protein and is implicated in ribosomal biogenesis.
Immunogen
This protein G purified monoclonal antibody was produced by repeated immunizations with a synthetic peptide corresponding to amino acid residues 54-75 of mouse p19Arf protein.
Immunogen Type
Peptide
Storage Condition
Store vial at -20° C prior to opening.  Aliquot contents and freeze at -20° C or below for extended storage.  Avoid cycles of freezing and thawing.  Centrifuge product if not completely clear after standing at room temperature.  This product is stable for several weeks at 4° C as an undiluted liquid.  Dilute only prior to immediate use.
Application Note
This antibody has been tested for use in ELISA, western blotting, immunofluorescence microscopy, and immunoprecipitation. Specific conditions for western blotting reactivity should be optimized by the end user.  Expect a band approximately 19 kDa in size corresponding to mouse p19Arf by western blotting in the appropriate cell line or lysate.
Purity/Specifity
This is a Protein G purified antibody directed against mouse p19Arf protein.  No reactivity is observed against human or hamster homologues.  The epitope was putatively mapped to amino acids 62-75 of mouse p19Arf.  BLAST analysis indicates that no significant sequence homology exists for this sequence with p19Arf homologues from other sources.   The homologue from rat only shows 11 of 14 identities (78% homology).  No additional information is available for reactivity with p19Arf protein from other sources.
Disclaimer Note-General
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.
General Reference
Quelle,D.E., Zindy,F., Ashmun,R.A. and Sherr,C.J. (1995) Alternative reading frames of the INK4a tumor suppressor gene encode two unrelated proteins capable of inducing cell cycle arrest. Cell 83 (6), 993-1000. Serrano, M., Hannon, G. J. & Beach, D. (1993) A new regulatory motif in cell-cycle control causing specific inhibition of cyclin D/CDK4. Nature 366, 704–707. Kamijo, T., Zindy, F., Roussel, M. F., Quelle, D. E., Downing, J. R., Ashmun, R. A., Grosveld, G. & Sherr, C. J. (1997) Tumor suppression at the mouse INK4a locus mediated by the alternative reading frame product p19ARF. Cell 91, 649–659. Lowe, S. W. & Sherr, C. J. (2003) Tumor suppression by Ink4a–Arf: progress and puzzles. Curr. Opin. Genet. Dev. 13, 77–83. Kamijo, T., van de Kamp, E., Chong, M. J., Zindy, F., Diehl, A. J., Sherr, C. J. & McKinnon, P. (1999) Loss of the ARF tumor suppressor reverses premature replicative arrest but not radiation hypersensitivity arising from disabled atm function. Cancer Res. 59, 2464–2469. Eischen, C. M., Weber, J. D., Roussel, M. F., Sherr, C. J. & Cleveland, J. L. (1999) Disruption of the ARF-Mdm2-p53 tumor suppressor pathway in Myc-induced lymphomagenesis. Genes Dev. 13, 2658–2669. Weber, J. D., Jeffers, J. R., Rehg, J. E., Randle, D. H., Lozano, G., Roussel, M. F., Sherr, C. J. & Zambetti, G. P. (2000) p53-independent functions of the p19(ARF) tumor suppressor. Genes Dev. 14, 2358–2365. Sugimoto, M., Kuo, M. L., Roussel, M. F. & Sherr, C. J. (2003) Nucleolar Arf tumor suppressor inhibits ribosomal RNA processing. Mol. Cell 11, 415–424.
Specific Reference
Bertwistle D, Zindy F, Sherr CJ, Roussel MF. (2004) Monoclonal antibodies to the mouse p19(Arf) tumor suppressor protein. Hybrid Hybridomics 23(5):293-300.
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Product Type
Primary Antibodies;
Applications
ELISA, WB
Reacts With
human
Catalog Number
100-401-224

Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, IP, ChIP
Reacts With
human
Catalog Number
200-301-174

Product Type
Primary Antibodies;
Applications
ELISA, WB, IP
Reacts With
human, mouse, hamster.
Catalog Number
200-501-892

Product Type
Secondary Antibodies;
Applications
ELISA, WB, IHC
Reacts With
Rat
Catalog Number
612-103-120

Product Label

 

Description
Unconjugated
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry