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PC116 Anti-ATM (Ab-3) (819-844) Rabbit pAb

Anti-ATM (Ab-3) (819-844), rabbit polyclonal, recognizes the ~350 kDa ATM protein in Daudi and HeLa cells. It is validated for Western blotting and immunoprecipitation.

Anti-ATM (Ab-3) (819-844) Rabbit pAb: Ficha de datos de seguridad (MSDS o SDS), certificado de análisis y de calidad (CoA y CoQ), expedientes, folletos y otros documentos disponibles.

Sinónimos: Anti-Ataxia Telangiectasia

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      Descripción

      Replacement Information

      Tabla espec. clave

      Species ReactivityHostAntibody Type
      HRbPolyclonal Antibody
      Description
      OverviewRecognizes the ~350 kDa ATM protein in Daudi and HeLa cells.
      Catalogue NumberPC116
      Brand Family Calbiochem®
      SynonymsAnti-Ataxia Telangiectasia
      References
      ReferencesFriedberg, E.C., et al. 1995. Amer. Soc. of Microbiolgy (meeting report), Wash. D.C.
      Meyn, S.M. 1995. Cancer Res. 55, 5991.
      Paules, R.S., et al. 1995. Cancer Res. 55, 1763.
      Savitsky, K., et al. 1995. Science 268, 1749.
      Savitsky, K., et al. 1995. Hum. Molec. Genet. 4, 2025.
      Zakian, V., 1995. Cell 82, 685.
      Beamish, H., et al. 1993. Rad. Res. 138, 130.
      Kastan, M.B., et al. 1992. Cell 71, 587.
      Painter, R.B. and Young, B.R. 1980. Proc. Natl. Acad. Sci. USA 77, 7315.
      Product Information
      FormLiquid
      FormulationIn 50 mM sodium phosphate buffer, 0.2% gelatin.
      Negative controlGM02052A cells
      Positive controlDaudi or HeLa cells
      Preservative≤0.1% sodium azide
      Quality LevelMQ100
      Applications
      Key Applications Immunoblotting (Western Blotting)
      Immunoprecipitation
      Application NotesImmunoblotting (2 µg/ml, see comments)
      Immunoprecipitation (2 µg)
      Application CommentsFor immunoblotting use 100-150 µg cell lysate on a 5% acrylamide gel (SDS/PAGE) and transferred to nitrocellulose using semi-dry transfer at 9V constant voltage for 2 h. Detection of antibody/antigen complexes is done using HRP conjugated goat anti-rabbit IgG at 25 ng/ml (Cat. No. DC03L).

      1. Anti-ATM (Ab-3) Rabbit pAb immunoblots and immunoprecipitates a 350 kDa protein in lysates of normal and transformed cells or tumor lines derived from individuals homozygous wild type for the ATM gene; whereas in cells derived from patients with Ataxia-Telangiectasia and carrying homozygous inactivating mutations in ATM, no 350 kDa protein is detected.
      2. Anti-ATM (Ab-3) Rabbit pAb may non-specifically detect smaller molecular weight proteins present in both ATM mutant and wild type cells. Careful titering of primary and secondary antibodies is recommended.
      3. Immunoblotting of p350ATM requires loading 100-150 µg cell lysate on low percentage acrylamide gels (5%) (SDS/PAGE) with electrophoresis performed until the 200 kDa molecular weight marker has migrated halfway through the gel. Semi-dry electrophoretic transfer is for 2 h at 9V constant voltage. Tank transfer is overnight at 40 V constant voltage.
      4. To transfer the gel for blotting, lay a dry piece of Whatman 3MM Chromatography paper over the wet gel. Carefully peel the 3MM paper and gel off the glass plate and immerse, gel side up, in transfer buffer until 3MM paper is thoroughly wet. Remove bubbles by rolling a pipette across the surface of the gel.
      5. To confirm detection of p350ATM, a cell line carrying a truncating mutation in ATM should be used as a negative control. Cell lines from A-T patients which show no detectable band at 350 kDa, can be obtained from the National Institute of General Medical Sciences Human Genetic Mutant Cell Repository at the Coriell Institute for Medical Research, 401 Haddon Avenue, Camden, NJ 08103.
      6. For immunoprecipitations, prepare nuclear lysates as described. Immunoprecipitate p350ATM using 2 µg of Anti-ATM (Ab-3) Rabbit pAb and Protein A-Agarose (Cat. No. IP06). Detection can be after metabolic labeling with 35S methionine followed by autoradiography, or alternatively, immunoprecipitated proteins can be displayed on 5% SDS/PAGE, transferred to nitrocellulose and then blotted as above using Anti-ATM (Ab-3).
      Biological Information
      Immunogena synthetic peptide (CKSLASFIKKPFDRGEVESMEDDTNG) corresponding to amino acids 819-844 of human ATM
      ImmunogenHuman
      HostRabbit
      IsotypeIgG
      Species Reactivity
      • Human
      Antibody TypePolyclonal Antibody
      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 Standard Handling
      Storage +2°C to +8°C
      Do not freeze Yes
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Número de referencia GTIN
      PC116 0

      Documentation

      Anti-ATM (Ab-3) (819-844) Rabbit pAb Ficha datos de seguridad (MSDS)

      Título

      Ficha técnica de seguridad del material (MSDS) 

      Anti-ATM (Ab-3) (819-844) Rabbit pAb Certificados de análisis

      CargoNúmero de lote
      PC116

      Referencias bibliográficas

      Visión general referencias
      Friedberg, E.C., et al. 1995. Amer. Soc. of Microbiolgy (meeting report), Wash. D.C.
      Meyn, S.M. 1995. Cancer Res. 55, 5991.
      Paules, R.S., et al. 1995. Cancer Res. 55, 1763.
      Savitsky, K., et al. 1995. Science 268, 1749.
      Savitsky, K., et al. 1995. Hum. Molec. Genet. 4, 2025.
      Zakian, V., 1995. Cell 82, 685.
      Beamish, H., et al. 1993. Rad. Res. 138, 130.
      Kastan, M.B., et al. 1992. Cell 71, 587.
      Painter, R.B. and Young, B.R. 1980. Proc. Natl. Acad. Sci. USA 77, 7315.
      Ficha técnica

      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.

      Revision21-November-2007 RFH
      SynonymsAnti-Ataxia Telangiectasia
      ApplicationImmunoblotting (2 µg/ml, see comments)
      Immunoprecipitation (2 µg)
      DescriptionPurified rabbit polyclonal antibody. Recognizes the ~350 kDa ATM protein.
      BackgroundAtaxia telangiectasia (AT) is an autosomal recessive disease characterized by cerebellar degeneration, an uneven gait (ataxia), neuromotor deterioration, the appearance of dilated blood vessels (telangiectasia) in the conjunctiva of the eyes and skin, and a large increase in cancer susceptibility. Cells of AT patients have increased sensitivity to ionizing radiation, defects in G1 and G2 of the cell cycle, and are unable to effect a normal p53-dependent G1 arrest due to failure to properly induce WAF1. Identification of the gene in AT patients showed that the disease is caused by a mutation of a single gene called ATM. Cloning of the ATM cDNA indicates that it contains a large open reading frame and encodes a putative protein of 3056 amino acids. The C-terminal region has extensive homology to the catalytic domains of phosphatidylinositol 3-kinases (PI3 kinases). A family of ATM-related genes has been identifed suggesting a role for these proteins in signal transduction and cell cycle regulation. ATM may be a key regulator of p53-mediated apoptosis.
      HostRabbit
      Immunogen speciesHuman
      Immunogena synthetic peptide (CKSLASFIKKPFDRGEVESMEDDTNG) corresponding to amino acids 819-844 of human ATM
      IsotypeIgG
      Specieshuman
      Positive controlDaudi or HeLa cells
      Negative controlGM02052A cells
      FormLiquid
      FormulationIn 50 mM sodium phosphate buffer, 0.2% gelatin.
      Concentration Label Please refer to vial label for lot-specific concentration
      Preservative≤0.1% sodium azide
      CommentsFor immunoblotting use 100-150 µg cell lysate on a 5% acrylamide gel (SDS/PAGE) and transferred to nitrocellulose using semi-dry transfer at 9V constant voltage for 2 h. Detection of antibody/antigen complexes is done using HRP conjugated goat anti-rabbit IgG at 25 ng/ml (Cat. No. DC03L).

      1. Anti-ATM (Ab-3) Rabbit pAb immunoblots and immunoprecipitates a 350 kDa protein in lysates of normal and transformed cells or tumor lines derived from individuals homozygous wild type for the ATM gene; whereas in cells derived from patients with Ataxia-Telangiectasia and carrying homozygous inactivating mutations in ATM, no 350 kDa protein is detected.
      2. Anti-ATM (Ab-3) Rabbit pAb may non-specifically detect smaller molecular weight proteins present in both ATM mutant and wild type cells. Careful titering of primary and secondary antibodies is recommended.
      3. Immunoblotting of p350ATM requires loading 100-150 µg cell lysate on low percentage acrylamide gels (5%) (SDS/PAGE) with electrophoresis performed until the 200 kDa molecular weight marker has migrated halfway through the gel. Semi-dry electrophoretic transfer is for 2 h at 9V constant voltage. Tank transfer is overnight at 40 V constant voltage.
      4. To transfer the gel for blotting, lay a dry piece of Whatman 3MM Chromatography paper over the wet gel. Carefully peel the 3MM paper and gel off the glass plate and immerse, gel side up, in transfer buffer until 3MM paper is thoroughly wet. Remove bubbles by rolling a pipette across the surface of the gel.
      5. To confirm detection of p350ATM, a cell line carrying a truncating mutation in ATM should be used as a negative control. Cell lines from A-T patients which show no detectable band at 350 kDa, can be obtained from the National Institute of General Medical Sciences Human Genetic Mutant Cell Repository at the Coriell Institute for Medical Research, 401 Haddon Avenue, Camden, NJ 08103.
      6. For immunoprecipitations, prepare nuclear lysates as described. Immunoprecipitate p350ATM using 2 µg of Anti-ATM (Ab-3) Rabbit pAb and Protein A-Agarose (Cat. No. IP06). Detection can be after metabolic labeling with 35S methionine followed by autoradiography, or alternatively, immunoprecipitated proteins can be displayed on 5% SDS/PAGE, transferred to nitrocellulose and then blotted as above using Anti-ATM (Ab-3).
      Storage +2°C to +8°C
      Do Not Freeze Yes
      Toxicity Standard Handling
      ReferencesFriedberg, E.C., et al. 1995. Amer. Soc. of Microbiolgy (meeting report), Wash. D.C.
      Meyn, S.M. 1995. Cancer Res. 55, 5991.
      Paules, R.S., et al. 1995. Cancer Res. 55, 1763.
      Savitsky, K., et al. 1995. Science 268, 1749.
      Savitsky, K., et al. 1995. Hum. Molec. Genet. 4, 2025.
      Zakian, V., 1995. Cell 82, 685.
      Beamish, H., et al. 1993. Rad. Res. 138, 130.
      Kastan, M.B., et al. 1992. Cell 71, 587.
      Painter, R.B. and Young, B.R. 1980. Proc. Natl. Acad. Sci. USA 77, 7315.