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OP45 Anti-DCC Mouse mAb (AF5)

Anti-DCC Mouse mAb (AF5): SDB (Sicherheitsdatenblätter), Analysenzertifikate und Qualitätszertifikate, Dossiers, Broschüren und andere verfügbare Dokumente.

Synonyme: Anti-Deleted Colorectal Carcinoma

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      Übersicht

      Replacement Information

      Key Spec Table

      Species ReactivityHostAntibody Type
      HMMonoclonal Antibody
      Description
      Overview

      This product has been discontinued.



      Recognizes the ~190 kDa DCC protein in IMR32 cells.
      Catalogue NumberOP45
      Brand Family Calbiochem®
      SynonymsAnti-Deleted Colorectal Carcinoma
      References
      ReferencesDunlop, M.G. 1992. Cancer Biol. 3, 131.
      Goyette, M.C., et al. 1992. Mol. Cell. Biol. 12, 1387.
      Hohne, M.W., et al. 1992. Cancer Res. 52, 2616.
      Huang, Y., et al. 1992. Cancer Res. 52, 6525.
      Kikuchi-Yanoshita, R., et al. 1992. Cancer Res. 52, 3801.
      Tanaka, K., et al. 1991. Nature 349, 340.
      Fearon, E.R., et al. 1990. Science 247, 49.
      Fearon, E.R., and Vogelstein, B. 1990. Cell 61, 759.
      Miyaki, M., et al. 1990. Cancer Res. 50, 7166.
      Vogelstein, B., et al. 1989. Science 244, 207.
      Product Information
      FormLiquid
      FormulationIn 0.05 M sodium phosphate buffer, 0.2% gelatin.
      Negative controlSW480 cells
      Positive controlIMR32 cells
      Preservative≤0.1% sodium azide
      Quality LevelMQ100
      Applications
      Key Applications Immunoblotting (Western Blotting)
      Immunofluorescence
      Immunoprecipitation
      Application NotesImmunoblotting (1-5 µg/ml)
      Immunofluorescence (1-5 µg/ml)
      Immunoprecipitation (1-2 µg/sample)
      Application CommentsIMMUNOBLOTTING PROCEDURE

      Anti-DCC (Ab-1) can be used to immunoblot proteins previously separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophoretically transferred onto nitrocellulose membranes. The proteins are reacted with the monoclonal antibody and visualized using an HRP conjugated goat anti-mouse antibody and chemiluminescent detection.

      Materials:

      1. Equipment

      • Electrophoresis apparatus
      • Rocker platform

      2. Solutions and Reagents

      • Anti-DCC (Ab-1), Cat. No. OP45
      • Anti-mouse IgG conjugated to horseradish peroxidase
      • Chemiluminescent detection reagents
      • 1x Laemmli Sample Buffer: 60 mM Tris-Cl, pH 6.8, 2% SDS, 10% glycerol, 100 mM DTT, 0.001% Bromophenol Blue
      • 7.5% polyacrylamide gel
      • Tris-buffered saline (TBS) pH 7.6: 20 mM Tris base, 137 mM NaCl, pH to 7.6 with HCl
      • TBS/0.1% Tween®-20 detergent (TBST)
      • 10% Non-fat Dry Milk in TBST
      • 5% Non-fat Dry Milk in TBST

      Procedure:

      1) Lyse cells in 1x Laemmli Sample Buffer. For adherent cells, lyse a 10 cm plate in 2 ml of sample buffer added directly to cells on the plate. Final protein concentration is approximately 1 mg/ml.
      2) Electrophorese 50-100 µg of total lysate protein on SDS/PAGE (7.5% polyacrylamide).
      3) Transfer the proteins onto a nitrocellulose membrane using an electroblotting apparatus.
      4) Block the membrane for 1 h in 10% non-fat dry milk in TBSTat room temperature with rocking. Use about 1 ml per cm2 of membrane.
      5) Incubate the membrane with 1 µg/ml of Anti-DCC (Ab-1) in 5% non-fat dry milk in TBST for 2 h at room temperature with rocking.
      6) Wash the membrane 3 times, 10 min each, in TBST at room temperature with rocking
      7) Incubate the membrane with 100 ng/ml of HRP conjugated goat anti-mouse IgG antibody in 5% non-fat dry milk in TBST at room temperature for 1 h.
      8) Wash the membrane 6 times, 10 min each, in TBST at room temperature with rocking.
      9) Develop the membrane using chemiluminescent detection reagents according to manufacturers instructions.
      10) Expose the membrane to film for 10 min. Adjust exposure times (from 30 s to 24 h) as necessary.
      Biological Information
      Immunogenthe extracellular domain of human DCC
      ImmunogenHuman
      CloneAF5
      HostMouse
      IsotypeIgG₁
      Species Reactivity
      • Human
      Antibody TypeMonoclonal 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
      Bestellnummer GTIN
      OP45 0

      Documentation

      Anti-DCC Mouse mAb (AF5) SDB

      Titel

      Sicherheitsdatenblatt (SDB) 

      Anti-DCC Mouse mAb (AF5) Analysenzertifikate

      TitelChargennummer
      OP45

      Literatur

      Übersicht
      Dunlop, M.G. 1992. Cancer Biol. 3, 131.
      Goyette, M.C., et al. 1992. Mol. Cell. Biol. 12, 1387.
      Hohne, M.W., et al. 1992. Cancer Res. 52, 2616.
      Huang, Y., et al. 1992. Cancer Res. 52, 6525.
      Kikuchi-Yanoshita, R., et al. 1992. Cancer Res. 52, 3801.
      Tanaka, K., et al. 1991. Nature 349, 340.
      Fearon, E.R., et al. 1990. Science 247, 49.
      Fearon, E.R., and Vogelstein, B. 1990. Cell 61, 759.
      Miyaki, M., et al. 1990. Cancer Res. 50, 7166.
      Vogelstein, B., et al. 1989. Science 244, 207.
      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.

      Revision03-October-2007 RFH
      SynonymsAnti-Deleted Colorectal Carcinoma
      ApplicationImmunoblotting (1-5 µg/ml)
      Immunofluorescence (1-5 µg/ml)
      Immunoprecipitation (1-2 µg/sample)
      DescriptionPurified mouse monoclonal antibody. Recognizes the ~190 kDa DCC protein.
      BackgroundInvasive, metastatic colon cancer arises from pre-existing adenomas in both familial and sporadic cases and is characterized by the presence of multiple chromosomal alterations. In the progression from intramucosal carcinoma to invasive carcinoma, allelic loss on the long arm of chromosome 18 is frequently observed. Recently, a gene on 18q21.3 termed DCC, for deleted in colorectal carcinoma, has been isolated and shown to be deleted or mutated in both sporadic and inherited colon carcinoma whereas normal colonic tissue retained and expressed the gene. Furthermore, loss of DCC expression appears to accompany progression of adenomas to metastatic carcinoma. The inactivation of DCC suggests that it is a tumor suppressor gene. A functional role for DCC as a tumor suppressor gene is indicated by experiments demonstrating that introduction of a normal chromosome 18 into the human colon tumor cell lines COKFu or SW480.7 lacking DCC suppresses tumorigenicity. Analysis of the amino acid sequence deduced from cDNA clones indicates that DCC is a transmembrane glycoprotein consisting of 1447 amino acids with an extracellular domain having extensive similarity to neural cell adhesion molecules (NCAM), a single transmembrane segment, and a unique cytoplasmic domain. The high degree of similarity to NCAMs suggests that DCC is involved in cell-cell interactions essential to the differentiated state of the colonic epithelium. DCC expression has been observed in all tissues examined except liver with the highest level of expression in brain tissue. Alterations in DCC expression have also been observed in pancreatic adenocarcinoma and esophogeal carcinoma. However, DCC mRNA is expressed at very low levels, requiring reverse transcription followed by PCR amplification for unambiguous detection.
      HostMouse
      Immunogen speciesHuman
      Immunogenthe extracellular domain of human DCC
      CloneAF5
      IsotypeIgG₁
      Specieshuman
      Positive controlIMR32 cells
      Negative controlSW480 cells
      FormLiquid
      FormulationIn 0.05 M sodium phosphate buffer, 0.2% gelatin.
      Concentration Label Please refer to vial label for lot-specific concentration
      Preservative≤0.1% sodium azide
      CommentsIMMUNOBLOTTING PROCEDURE

      Anti-DCC (Ab-1) can be used to immunoblot proteins previously separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophoretically transferred onto nitrocellulose membranes. The proteins are reacted with the monoclonal antibody and visualized using an HRP conjugated goat anti-mouse antibody and chemiluminescent detection.

      Materials:

      1. Equipment

      • Electrophoresis apparatus
      • Rocker platform

      2. Solutions and Reagents

      • Anti-DCC (Ab-1), Cat. No. OP45
      • Anti-mouse IgG conjugated to horseradish peroxidase
      • Chemiluminescent detection reagents
      • 1x Laemmli Sample Buffer: 60 mM Tris-Cl, pH 6.8, 2% SDS, 10% glycerol, 100 mM DTT, 0.001% Bromophenol Blue
      • 7.5% polyacrylamide gel
      • Tris-buffered saline (TBS) pH 7.6: 20 mM Tris base, 137 mM NaCl, pH to 7.6 with HCl
      • TBS/0.1% Tween®-20 detergent (TBST)
      • 10% Non-fat Dry Milk in TBST
      • 5% Non-fat Dry Milk in TBST

      Procedure:

      1) Lyse cells in 1x Laemmli Sample Buffer. For adherent cells, lyse a 10 cm plate in 2 ml of sample buffer added directly to cells on the plate. Final protein concentration is approximately 1 mg/ml.
      2) Electrophorese 50-100 µg of total lysate protein on SDS/PAGE (7.5% polyacrylamide).
      3) Transfer the proteins onto a nitrocellulose membrane using an electroblotting apparatus.
      4) Block the membrane for 1 h in 10% non-fat dry milk in TBSTat room temperature with rocking. Use about 1 ml per cm2 of membrane.
      5) Incubate the membrane with 1 µg/ml of Anti-DCC (Ab-1) in 5% non-fat dry milk in TBST for 2 h at room temperature with rocking.
      6) Wash the membrane 3 times, 10 min each, in TBST at room temperature with rocking
      7) Incubate the membrane with 100 ng/ml of HRP conjugated goat anti-mouse IgG antibody in 5% non-fat dry milk in TBST at room temperature for 1 h.
      8) Wash the membrane 6 times, 10 min each, in TBST at room temperature with rocking.
      9) Develop the membrane using chemiluminescent detection reagents according to manufacturers instructions.
      10) Expose the membrane to film for 10 min. Adjust exposure times (from 30 s to 24 h) as necessary.
      Storage +2°C to +8°C
      Do Not Freeze Yes
      Toxicity Standard Handling
      ReferencesDunlop, M.G. 1992. Cancer Biol. 3, 131.
      Goyette, M.C., et al. 1992. Mol. Cell. Biol. 12, 1387.
      Hohne, M.W., et al. 1992. Cancer Res. 52, 2616.
      Huang, Y., et al. 1992. Cancer Res. 52, 6525.
      Kikuchi-Yanoshita, R., et al. 1992. Cancer Res. 52, 3801.
      Tanaka, K., et al. 1991. Nature 349, 340.
      Fearon, E.R., et al. 1990. Science 247, 49.
      Fearon, E.R., and Vogelstein, B. 1990. Cell 61, 759.
      Miyaki, M., et al. 1990. Cancer Res. 50, 7166.
      Vogelstein, B., et al. 1989. Science 244, 207.