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AM30 Anti-PARP-1 (Ab-2) Mouse mAb (C-2-10)

Anti-PARP-1 (Ab-2) Mouse mAb (C-2-10) : FDS (Fiches de données de sécurité), certificats d’analyse (CoA) et de qualité (CoQ), dossiers, brochures et autres documents disponibles.

Synonymes: Anti-Poly(ADP-Ribose) Polymerase

AM30
  
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      Description
      Overview

      This product has been discontinued.





      Recognizes the ~116 kDa full-length PARP protein and the ~85 kDa cleaved fragment of PARP in etopside-treated HL-60 cells.
    • Antibody Target Gene Symbol: PARP1
    • Target Synonym: 5830444G22Rik, Adprp, Adprp1, ADPRT, ADPRT1, AI893648, C80510, MGC93658, PADPRT, pADPRT-1, PARP, PARS, POLY(ADP-RIBOSE) POLYMERASE 1, PPOL, sPARP-1
    • Entrez Gene Name: poly (ADP-ribose) polymerase 1
    • Hu Entrez ID: 142 142 (Related Antibodies: PC100, AM68, 512739, 512738, 512737, and 512734)
    • Mu Entrez ID: 11545
    • Rat Entrez ID: 25591
      		•
      Catalogue NumberAM30
      Brand Family Calbiochem®
      SynonymsAnti-Poly(ADP-Ribose) Polymerase
      Application Data
      Detection of human PARP-1 by immunoblotting. Samples: Whole cell lysate from HL60 cells left untreated (lane 1) and treated with etoposide (lane 2). Primary antibody: Anti-PARP-1 (Ab-2) Mouse mAb (C-2-10) (Cat. No. AM30) (1 µg/ml). Detection: chemiluminescence.
      References
      ReferencesLeist, M., et al. 1997. Biochem. Biophys. Res. Commun. 233, 518.
      Posmantur, R., et al. 1997. J. Neurochem. 68, 2328.
      Rosen, A., and Casciola-Rosen, L. 1997. J. Cell. Biochem. 64, 50.
      Rosenthal, D.S., et al. 1997. Exp. Cell. Res. 232, 313.
      Whitacre, C.M. and Berger, N.A. 1997. Cancer Res. 57, 2157.
      Shah, G.M., et al. 1996. Biochem. Biophys. Res. Commun. 229,838.
      Yoshida, S., and Simbulan, C.M. 1994. Mol. Cell Biochem. 138, 39.
      Satoh, M.S., and Lindahl, T. 1992. Nature 356, 356.
      Lamarre, D. et al. 1988. Biochem Biophys Acta 95.
      Berger, N.A. 1985. Radiat. Res. 101, 4.
      Product Information
      FormLiquid
      FormulationIn PBS, 0.2% BSA.
      Positive controlHL60 cells treated with etoposide
      Preservative≤0.1% sodium azide
      Applications
      Application ReferencesImmunoblotting Hobom, U. and Dobbelstein, M. 2004. J. Virol. 78, 7685. Ungefroren, H., et al. 2001. J. Cell Sci. 114, 2735. Grandgirard, D., et al. 1998. EMBO J. 17,1268.
      Key Applications Immunoblotting (Western Blotting)
      Immunofluorescence
      Not Frozen Sections
      Not Paraffin Sections
      Application NotesFrozen Sections (not recommended)
      Immunoblotting (1 µg/ml, see application references)
      Immunofluorescence (2 µg/ml)
      Paraffin Sections (not recommended)
      Application CommentsRecognizes the C-terminus of the PARP DNA-binding domain. This antibody may be used to stain cytospin preparations of HL-60 cells treated with etoposide. Many cells can be used as a positive control, including HL-60 cells treated with etoposide and MCF-7 cells treated with dexamethasone. Untreated cells should express only full-length PARP while treated cells will express cleaved and full-length PARP. For immunoblotting, sonication and urea are necessary to break up PARP/DNA interactions. To do this, resuspend cells in 62.5 mM Tris/HCl, pH 6.8, 6 M urea, 10% glycerol, 2% SDS, 0.00125% bromophenol blue and 5% β-mercaptoethanol, then sonicate for 15 s on full power and incubate at 65°C for 15 min. Antibody should be titrated for optimal staining in individual systems.

      Immunoblotting with Anti-PARP

      1. HL-60 cells are prepared from exponentially growing cells either induced for apoptosis with etoposide (Cat. No. 341205) or uninduced. Cells are washed once with PBS, suspended at ~4 x 106 cells/ml in sample buffer (62.5 mM Tris-HCl, pH 6.8, 6 M urea, 10% glycerol, 2% SDS, 0.00125% bromophenol blue, 5% β-mercaptoethanol. Cells are sonicated for 15 s and incubated at 65°C for 15 min.
      Note: The purpose of the urea and sonication step is to effectively dissociate PARP/DNA interactions. For preparing PARP electrophoresis samples from tissue, see Shah, G.M., et al. 1995. Anal. Biochem. 227, 1 or Simonin, F., et al. 1991. Anal. Biochem. 195, 226.

      2. Run SDS-PAGE using 20 µl of control or induced HL-60 extract.

      3. Transfer proteins to nitrocellulose blotting membrane.

      4. Block with 5% non-fat dry milk in TBST (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.1% Tween® 20 detergent) for 1 h at room temperature.

      5. Incubate nitrocellulose with the appropriate dilution of anti-PARP antibody in TBST with 5% non-fat dry milk for 2.5 h at room temperature.

      6. Wash membrane 3 x with TBST, 10 min/wash.

      7. Incubate nitrocellulose with secondary antibody in TBST with 5% milk, using either anti-mouse or anti-rabbit alkaline phosphatase conjugate, as appropriate, for 1 h at room temperature.

      8. Wash membrane 3x in TBST, 10 min per wash. Rinse briefly with TBS (TBST without the Tween® 20 detergent).

      9. Incubate with BCIP/NBT color development reagent for ~5 min until bands reach desired intensity. Rinse with TBS plus 20 mM EDTA to stop color development. Note: color development times may vary.
      Biological Information
      Immunogenpurified PARP (poly(ADP-ribose) polymerase) from calf thymus
      ImmunogenBovine
      CloneC-2-10
      HostMouse
      IsotypeIgG₁
      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 Dry Ice Only
      Toxicity Standard Handling
      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
      Référence GTIN
      AM30 0

      Documentation

      Anti-PARP-1 (Ab-2) Mouse mAb (C-2-10) FDS

      Titre

      Fiche de données de sécurité des matériaux (FDS) 

      Anti-PARP-1 (Ab-2) Mouse mAb (C-2-10) Certificats d'analyse

      TitreNuméro de lot
      AM30

      Références bibliographiques

      Aperçu de la référence bibliographique
      Leist, M., et al. 1997. Biochem. Biophys. Res. Commun. 233, 518.
      Posmantur, R., et al. 1997. J. Neurochem. 68, 2328.
      Rosen, A., and Casciola-Rosen, L. 1997. J. Cell. Biochem. 64, 50.
      Rosenthal, D.S., et al. 1997. Exp. Cell. Res. 232, 313.
      Whitacre, C.M. and Berger, N.A. 1997. Cancer Res. 57, 2157.
      Shah, G.M., et al. 1996. Biochem. Biophys. Res. Commun. 229,838.
      Yoshida, S., and Simbulan, C.M. 1994. Mol. Cell Biochem. 138, 39.
      Satoh, M.S., and Lindahl, T. 1992. Nature 356, 356.
      Lamarre, D. et al. 1988. Biochem Biophys Acta 95.
      Berger, N.A. 1985. Radiat. Res. 101, 4.

      Brochure

      Titre
      Caspases and other Apoptosis Related Tools Brochure

      Citations

      Titre
    • Urs Hobom and Matthias Dobbelstein. (2004) E1B-55-kilodalton protein is not required to block p53-induced transcription during adenovirus infection. Journal of Virology 78, 7685-7697.
    • Cecile Tetaud, et al. (2003) Two Distinct Gb3/CD77 Signaling Pathways Leading to Apoptosis are Triggered by Anti-Gb3/CD77 mAb and Verotoxin-1. Journal of Biological Chemistry 278, 45200-45208.
    • Hendrik Ungefroren, et al. (2001) FAP-1 in pancreatic cancer cells: functional and mechanistic studies on its inhibitory role in CD95-mediated apoptosis. Journal of Cell Science 114, 2735-2746.
    • Brian L. Webb, Elsa Jimenez and G. Steven Martin. (2000) v-Src Generates a p53-Independent Apoptotic Signal. Molecular and Cellular Biology 20, 9271-9280.
      		•
      Fiche technique

      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.

      Revision18-March-2008 RFH
      SynonymsAnti-Poly(ADP-Ribose) Polymerase
      ApplicationFrozen Sections (not recommended)
      Immunoblotting (1 µg/ml, see application references)
      Immunofluorescence (2 µg/ml)
      Paraffin Sections (not recommended)
      Application Data
      Detection of human PARP-1 by immunoblotting. Samples: Whole cell lysate from HL60 cells left untreated (lane 1) and treated with etoposide (lane 2). Primary antibody: Anti-PARP-1 (Ab-2) Mouse mAb (C-2-10) (Cat. No. AM30) (1 µg/ml). Detection: chemiluminescence.
      DescriptionPurified mouse monoclonal antibody. Recognizes the ~116 kDa full length and the ~85 kDa cleaved forms of PARP.
      BackgroundPARP (Poly (ADP-ribose) polymerase [NAD+ ADP-ribosyltransferase; NAD+:poly(adenosine-diphosphate-D-ribosyl)-acceptor ADP-D-ribosyltransferase, EC 2.4.2.30] is a highly conserved nuclear enzyme present in higher eukaryotes. The enzyme is a Zn2+ dependent DNA binding protein that recognizes DNA strand breaks and is implicated in DNA repair and in the apoptotic response of cells. The critical functional domains of the protein are the DNA binding domain and the automodification domains. PARP functions by adding poly (ADP-ribose), in an ATP and NAD+ dependent manner, to various proteins in response to DNA damage and has been shown to directly interact with DNA polymerase α. As a marker for apoptosis, PARP cleavage has been variably shown to occur early in the response as a result of the activity of CPP32 (caspase-3, CASP3). PARP cleavage correlates well with chromatin condensation, and has been shown to be associated with the condensed chromatin in apoptotic cells, as a measure of apoptosis appearing as early as 3 h post apoptosis inducing event, and precedes the ability to detect actual DNA fragmentation. Although shown in many systems to be a marker for apoptosis it is not clear that PARP cleavage is required for apoptosis to occur based on results from studies in PARP -/- mice. Degradation of PARP has also been shown to occur during the process of necrosis however an entirely different pattern of cleavage products is seen. In necrosis, major cleavage products of 89 and 50 kDa as well as minor cleavage products of 40 and 35 kDa have been reported. This latter pattern differs from that seen during apoptosis when only a major fragment of ~89 kDa is observed.
      HostMouse
      Immunogen speciesBovine
      Immunogenpurified PARP (poly(ADP-ribose) polymerase) from calf thymus
      CloneC-2-10
      IsotypeIgG₁
      Speciesnot chicken, hamster, human, monkey, mouse, rat
      Positive controlHL60 cells treated with etoposide
      FormLiquid
      FormulationIn PBS, 0.2% BSA.
      Concentration Label Please refer to vial label for lot-specific concentration
      Preservative≤0.1% sodium azide
      CommentsRecognizes the C-terminus of the PARP DNA-binding domain. This antibody may be used to stain cytospin preparations of HL-60 cells treated with etoposide. Many cells can be used as a positive control, including HL-60 cells treated with etoposide and MCF-7 cells treated with dexamethasone. Untreated cells should express only full-length PARP while treated cells will express cleaved and full-length PARP. For immunoblotting, sonication and urea are necessary to break up PARP/DNA interactions. To do this, resuspend cells in 62.5 mM Tris/HCl, pH 6.8, 6 M urea, 10% glycerol, 2% SDS, 0.00125% bromophenol blue and 5% β-mercaptoethanol, then sonicate for 15 s on full power and incubate at 65°C for 15 min. Antibody should be titrated for optimal staining in individual systems.

      Immunoblotting with Anti-PARP

      1. HL-60 cells are prepared from exponentially growing cells either induced for apoptosis with etoposide (Cat. No. 341205) or uninduced. Cells are washed once with PBS, suspended at ~4 x 106 cells/ml in sample buffer (62.5 mM Tris-HCl, pH 6.8, 6 M urea, 10% glycerol, 2% SDS, 0.00125% bromophenol blue, 5% β-mercaptoethanol. Cells are sonicated for 15 s and incubated at 65°C for 15 min.
      Note: The purpose of the urea and sonication step is to effectively dissociate PARP/DNA interactions. For preparing PARP electrophoresis samples from tissue, see Shah, G.M., et al. 1995. Anal. Biochem. 227, 1 or Simonin, F., et al. 1991. Anal. Biochem. 195, 226.

      2. Run SDS-PAGE using 20 µl of control or induced HL-60 extract.

      3. Transfer proteins to nitrocellulose blotting membrane.

      4. Block with 5% non-fat dry milk in TBST (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.1% Tween® 20 detergent) for 1 h at room temperature.

      5. Incubate nitrocellulose with the appropriate dilution of anti-PARP antibody in TBST with 5% non-fat dry milk for 2.5 h at room temperature.

      6. Wash membrane 3 x with TBST, 10 min/wash.

      7. Incubate nitrocellulose with secondary antibody in TBST with 5% milk, using either anti-mouse or anti-rabbit alkaline phosphatase conjugate, as appropriate, for 1 h at room temperature.

      8. Wash membrane 3x in TBST, 10 min per wash. Rinse briefly with TBS (TBST without the Tween® 20 detergent).

      9. Incubate with BCIP/NBT color development reagent for ~5 min until bands reach desired intensity. Rinse with TBS plus 20 mM EDTA to stop color development. Note: color development times may vary.
      Storage Avoid freeze/thaw
      -20°C
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-20°C).
      Toxicity Standard Handling
      ReferencesLeist, M., et al. 1997. Biochem. Biophys. Res. Commun. 233, 518.
      Posmantur, R., et al. 1997. J. Neurochem. 68, 2328.
      Rosen, A., and Casciola-Rosen, L. 1997. J. Cell. Biochem. 64, 50.
      Rosenthal, D.S., et al. 1997. Exp. Cell. Res. 232, 313.
      Whitacre, C.M. and Berger, N.A. 1997. Cancer Res. 57, 2157.
      Shah, G.M., et al. 1996. Biochem. Biophys. Res. Commun. 229,838.
      Yoshida, S., and Simbulan, C.M. 1994. Mol. Cell Biochem. 138, 39.
      Satoh, M.S., and Lindahl, T. 1992. Nature 356, 356.
      Lamarre, D. et al. 1988. Biochem Biophys Acta 95.
      Berger, N.A. 1985. Radiat. Res. 101, 4.
      Citation
    • Urs Hobom and Matthias Dobbelstein. (2004) E1B-55-kilodalton protein is not required to block p53-induced transcription during adenovirus infection. Journal of Virology 78, 7685-7697.
    • Cecile Tetaud, et al. (2003) Two Distinct Gb3/CD77 Signaling Pathways Leading to Apoptosis are Triggered by Anti-Gb3/CD77 mAb and Verotoxin-1. Journal of Biological Chemistry 278, 45200-45208.
    • Hendrik Ungefroren, et al. (2001) FAP-1 in pancreatic cancer cells: functional and mechanistic studies on its inhibitory role in CD95-mediated apoptosis. Journal of Cell Science 114, 2735-2746.
    • Brian L. Webb, Elsa Jimenez and G. Steven Martin. (2000) v-Src Generates a p53-Independent Apoptotic Signal. Molecular and Cellular Biology 20, 9271-9280.
      		•
      Application referencesImmunoblotting Hobom, U. and Dobbelstein, M. 2004. J. Virol. 78, 7685. Ungefroren, H., et al. 2001. J. Cell Sci. 114, 2735. Grandgirard, D., et al. 1998. EMBO J. 17,1268.