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CB1012 Anti-p19ARF (54-75) Rat mAb (5-C3-1)

This Anti-p19ARF (54-75) Rat mAb (5-C3-1) is validated for use in Immunoblotting, IF, IP, Paraffin Sections for the detection of p19ARF (54-75).

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CB1012
  
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      Übersicht

      Replacement Information

      Key Spec Table

      Host
      R
      Description
      OverviewRecognizes the ~19 kDa p19ARF protein in whole cell lysates of mouse embryonic fibroblasts (MEF).

      This product has been discontinued.
      Catalogue NumberCB1012
      Brand Family Calbiochem®
      Application Data
      Detection of mouse p19ARF by immunoblotting. Samples: Whole cell lysate (30 μg) from MEF (lanes 1,3) and NIH3T3 (lanes 2,4) cells. Primary antibody: Anti-p19ARF (54-75) Rat mAb (12-A1-1) (Cat. No. CB1013) (3 μg/ml) (lanes 1,2) or Anti-p19ARF (54-75) Rat mAb (5-C3-1) (Cat. No. CB1012) (3 μg/ml) (lanes 3,4). Detection: chemiluminescence.

      A. Detection of mouse p19ARF by immunoblotting. Samples: Whole cell lysate (25 µg) from NIH3T3 cells that have deleted the Arf gene (lane 1) and wild type mouse embryo fibroblasts (MEFs) at passage 6, which express p19ARF (lane 2). Primary antibody: Anti-p19ARF (54-75) Rat mAb (5-C3-1) (Cat. No. CB1012) (3 µg/ml). Detection: chemiluminescence. B. Detection of mouse p19ARF by immunoprecipitation. Samples: Whole cell lysates (250 µg) from NIH3T3 cells that have deleted the Arf gene (lane 5) and NIH3T3 cells expressing p19ARF with a HA-tag (lanes 3,4). Antibodies for IP: Normal rat IgG (negative control; lane 3) and Anti-p19ARF (54-75) Rat mAb (5-C3-1) (Cat. No. CB1012). Conditions for immunoblotting: Samples: Whole cell lysate (25 µg, equivalent to 10% of above immunoprecipitations) from NIH3T3 cells that have deleted the Arf gene (lane 1), NIH3T3 cells expressing p19ARF with a HA-tag (lane 2), and immunoprecipitated proteins as indicated above. Primary antibody: Anti-HA. Detection: chemiluminescence. C. Detection of mouse p19ARF by immunocytochemistry. Samples: Wild type mouse embryo fibroblasts (MEF) at passage 5 (panels 1,2) and NIH3T3 cells, which have deleted the Arf gene (panels 3,4), fixed in methanol/acetone. Primary antibody: Anti-p19ARF (54-75) Rat mAb (5-C3-1) (Cat. No. CB1012). Detection: Anti-Rat IgG secondary antibody, fluorescent conjugate (panels 1,3) and counterstained with Hoechst dye (panels 2,4). D. Detection of mouse p19ARF by staining paraffin sections. Sample: Wild-type mouse testis. Primary antibody: Anti-p19ARF (54-75) Rat mAb (5-C3-1) (Cat. No. CB1012) (green). Detection: fluorescence with DAPI counterstain (blue).
      References
      ReferencesDe Stanchina, E., et al. 1998. Genes Dev. 12, 2434.
      Larsen, C.J., 1998. Bull. Cancer 85, 304.
      Radfar, A., et al. 1998. Proc. Natl. Acad. Sci. USA 95, 13194.
      Zindy, F., et al. 1998. Genes Dev. 12, 2424.
      Product Information
      FormLiquid
      FormulationIn PBS.
      Positive controlMouse embryonic fibroblasts (MEF)
      Preservative≤0.1% sodium azide
      Applications
      Application ReferencesOriginal Clone, Immunoblotting, Immunofluorescence, Immunoprecipitation Bertwistle, D., et al. 2004. Hybrid. Hybridomics 23, 293.
      Key Applications Immunoblotting (Western Blotting)
      Immunofluorescence
      Immunoprecipitation
      Paraffin Sections
      Application NotesImmunoblotting (3 µg/ml, see application references)
      Immunofluorescence (see application references)
      Immunoprecipitation (see application references)
      Paraffin Sections (see application data)
      Application CommentsRecognizes in vitro translated murine p19ARF and a 19 kDa band in murine CTLL2 cells. p19ARF staining can be seen in subnuclear structures (presumably nucleoli) in cells at the periphery of the testicular tubules. Staining is not detected in sections from a testis from an Arf null mouse (not shown). Antibody should be titrated for optimal results in individual systems.
      Biological Information
      Immunogena synthetic peptide corresponding to amino acids 54-75 of mouse p19ARF
      ImmunogenMouse
      Clone5-C3-1
      HostRat
      IsotypeIgG2b
      Concentration Label Please refer to vial label for lot-specific concentration
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Blue Ice Only
      Toxicity Irritant
      Storage -20°C
      Avoid freeze/thaw Avoid freeze/thaw
      Do not freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-20°C).
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Bestellnummer GTIN
      CB1012 0

      Documentation

      Anti-p19ARF (54-75) Rat mAb (5-C3-1) SDB

      Titel

      Sicherheitsdatenblatt (SDB) 

      Anti-p19ARF (54-75) Rat mAb (5-C3-1) Analysenzertifikate

      TitelChargennummer
      CB1012

      Literatur

      Übersicht
      De Stanchina, E., et al. 1998. Genes Dev. 12, 2434.
      Larsen, C.J., 1998. Bull. Cancer 85, 304.
      Radfar, A., et al. 1998. Proc. Natl. Acad. Sci. USA 95, 13194.
      Zindy, F., et al. 1998. Genes Dev. 12, 2424.
      Datenblatt

      Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

      Revision20-August-2007 RFH
      ApplicationImmunoblotting (3 µg/ml, see application references)
      Immunofluorescence (see application references)
      Immunoprecipitation (see application references)
      Paraffin Sections (see application data)
      Application Data
      Detection of mouse p19ARF by immunoblotting. Samples: Whole cell lysate (30 μg) from MEF (lanes 1,3) and NIH3T3 (lanes 2,4) cells. Primary antibody: Anti-p19ARF (54-75) Rat mAb (12-A1-1) (Cat. No. CB1013) (3 μg/ml) (lanes 1,2) or Anti-p19ARF (54-75) Rat mAb (5-C3-1) (Cat. No. CB1012) (3 μg/ml) (lanes 3,4). Detection: chemiluminescence.

      A. Detection of mouse p19ARF by immunoblotting. Samples: Whole cell lysate (25 µg) from NIH3T3 cells that have deleted the Arf gene (lane 1) and wild type mouse embryo fibroblasts (MEFs) at passage 6, which express p19ARF (lane 2). Primary antibody: Anti-p19ARF (54-75) Rat mAb (5-C3-1) (Cat. No. CB1012) (3 µg/ml). Detection: chemiluminescence. B. Detection of mouse p19ARF by immunoprecipitation. Samples: Whole cell lysates (250 µg) from NIH3T3 cells that have deleted the Arf gene (lane 5) and NIH3T3 cells expressing p19ARF with a HA-tag (lanes 3,4). Antibodies for IP: Normal rat IgG (negative control; lane 3) and Anti-p19ARF (54-75) Rat mAb (5-C3-1) (Cat. No. CB1012). Conditions for immunoblotting: Samples: Whole cell lysate (25 µg, equivalent to 10% of above immunoprecipitations) from NIH3T3 cells that have deleted the Arf gene (lane 1), NIH3T3 cells expressing p19ARF with a HA-tag (lane 2), and immunoprecipitated proteins as indicated above. Primary antibody: Anti-HA. Detection: chemiluminescence. C. Detection of mouse p19ARF by immunocytochemistry. Samples: Wild type mouse embryo fibroblasts (MEF) at passage 5 (panels 1,2) and NIH3T3 cells, which have deleted the Arf gene (panels 3,4), fixed in methanol/acetone. Primary antibody: Anti-p19ARF (54-75) Rat mAb (5-C3-1) (Cat. No. CB1012). Detection: Anti-Rat IgG secondary antibody, fluorescent conjugate (panels 1,3) and counterstained with Hoechst dye (panels 2,4). D. Detection of mouse p19ARF by staining paraffin sections. Sample: Wild-type mouse testis. Primary antibody: Anti-p19ARF (54-75) Rat mAb (5-C3-1) (Cat. No. CB1012) (green). Detection: fluorescence with DAPI counterstain (blue).
      DescriptionProtein G purified rat monoclonal antibody derived by immunizing rats with the specified immunogen and fusing splenocytes with Y3-Ag 1.2.3 rat myeloma cells. Recognizes the ~19 kDa p19ARF protein.
      BackgroundIn humans, the multiple tumor suppressor 1 locus (MTS1) encodes two unrelated genes, p16INK4a and p19ARF. Although both act as cell proliferation inhibitors, their mechanisms of action are different. p19ARF works as part of a p53-dependent pathway to counter uncontrolled proliferation and oncogenic signals. Mice lacking the p19ARF gene rapidly develop a broad spectrum of tumors. This suggests that p19ARF is an important tumor suppressor. In addition, p19ARF helps prevent transforming signals from various oncoproteins via the p53 regulatory loop. These studies and others demonstrate the importance of p19ARF in cell cycle control and tumor suppression.
      HostRat
      Immunogen speciesMouse
      Immunogena synthetic peptide corresponding to amino acids 54-75 of mouse p19ARF
      Clone5-C3-1
      IsotypeIgG2b
      Speciesmouse
      Positive controlMouse embryonic fibroblasts (MEF)
      FormLiquid
      FormulationIn PBS.
      Concentration Label Please refer to vial label for lot-specific concentration
      Preservative≤0.1% sodium azide
      CommentsRecognizes in vitro translated murine p19ARF and a 19 kDa band in murine CTLL2 cells. p19ARF staining can be seen in subnuclear structures (presumably nucleoli) in cells at the periphery of the testicular tubules. Staining is not detected in sections from a testis from an Arf null mouse (not shown). Antibody should be titrated for optimal results in individual systems.
      Storage Avoid freeze/thaw
      -20°C
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-20°C).
      Toxicity Irritant
      ReferencesDe Stanchina, E., et al. 1998. Genes Dev. 12, 2434.
      Larsen, C.J., 1998. Bull. Cancer 85, 304.
      Radfar, A., et al. 1998. Proc. Natl. Acad. Sci. USA 95, 13194.
      Zindy, F., et al. 1998. Genes Dev. 12, 2424.
      Application referencesOriginal Clone, Immunoblotting, Immunofluorescence, Immunoprecipitation Bertwistle, D., et al. 2004. Hybrid. Hybridomics 23, 293.