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OP62 Anti-APC (Ab-5) Mouse mAb (CF11)

Anti-APC (Ab-5) Mouse mAb (CF11) MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

Synonyms: Anti-Adenomatous Polyposis Coli

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      Overview

      Replacement Information

      Key Spec Table

      Species ReactivityHostAntibody Type
      H, MMMonoclonal Antibody
      Description
      Overview

      This product has been discontinued.



      Recognizes all full-length (p300) and truncated (p147) forms of APC.

    • Antibody Target Gene Symbol: APC
    • Target Synonym: AI047805, Apc7, AU020952, AW124434, BTPS2, DP2, DP2.5, DP3, Familial adenomatous polyposis, FAP, GS, Min, RATAPC
    • Entrez Gene Name: adenomatous polyposis coli
    • Hu Entrez ID: 324 (Related Antibodies: OP80, ST1150, OP47L, OP44)
    • Mu Entrez ID: 11789
    • Rat Entrez ID: 24205

      		•
      Catalogue NumberOP62
      Brand Family Calbiochem®
      SynonymsAnti-Adenomatous Polyposis Coli
      References
      ReferencesSmith, K.J., et al. 1993. Proc. Natl. Acad. Sci. USA 90, 2846.
      Su, L.-K., et al. 1993. Can. Res. 53, 2728.
      Boynton, R.F., et al. 1992. Proc. Natl. Acad. Sci. USA 89, 3385.
      D'Amico, D., et al. 1992. Cancer Res. 52, 1996.
      Fearon, E.R., and Jones, P.A. 1992. FASEB J. 6, 2783.
      Miyoshi, Y., et al. 1992. Proc. Natl. Acad. Sci. USA 89, 4452.
      Powell, S.M., et al. 1992. Nature 359, 235.
      Groden, J., et al. 1991. Cell 66, 589.
      Kinzler, K.W., et al. 1991. Science 253, 661.
      Nishisho, I., et al. 1991. Science 253, 665.
      Product Information
      FormLiquid
      FormulationIn 50 mM sodium phosphate buffer, 0.2% gelatin.
      Positive controlHEK293, A-431, HCT116, or HT-1080 cells (full-length APC) or SW480 cells (truncated APC)
      Preservative≤0.1% sodium azide
      Applications
      Application ReferencesImmunoprecipitation Su, L.-K., et al. 1993 Can. Res. 53, 2728.
      Key Applications Immunoprecipitation
      Application NotesImmunoprecipitation (see comments and application references)
      Application CommentsUsing this antibody various truncated and full lenghth forms of the APC protein have been observed in tumor cell lines and in lymphocytes obtained from FAP patients by immunoprecipiation. APC expression and turnover are very low making metabolic labelling difficult. APC immunocomplexes can be immunoprecipitated using Anti-APC (Ab-5) and detected in a immunoblot using Anti-APC (Ab-1), Cat. No. OP44 or Anti-APC (Ab-2), Cat. No. OP47L.

      IMMUNOPRECIPITATION PROCEDURE

      Can be used to immunoprecipitate full length and truncated APC protein APC. Precipitated APC proteins should then be visualized using APC (Ab-1) or APC (Ab-2) and the procedure provided with those products. APC expression and turnover are very low, therefore, immunoprecipitation of metabolically labeled lysates is not recommended.

      Materials:

      Equipment
      • Rocking Platform

      Solutions and Reagents:
      • Anti-APC (Ab-5), Mouse mAb (CF11) Cat. No. OP62
      • Protein A Sepharose swollen 1:1 in Wash Buffer
      • Normal Mouse IgG
      • Lysis Buffer: 100 mM NaCl, 50 mM Tris-HCl pH 7.5, 0.5% NP-40 Alternative, 0.5 µg/ml Leupeptin, 1.0 µg/ml Pepstatin, 0.2 mM PMSF
      • Wash Buffer: 100 mM NaCl, 50 mM Tris-HCl pH 7.5, 0.5% NP-40 Alternative
      • 2X Laemmli Sample Buffer: 120 mM Tris-HCl pH 6.8, 4% SDS, 20% glycerol, 200 mM DTT, 0.01% Bromophenol Blue

      Procedure:
      1) Lyse cells in lysis buffer. For adherent cells, lyse an 80-90% confluent 10 cm plate in 1.5 ml of buffer. Incubate at 4°C for 30 min with rocking. This will make enough lysate for 5 immunoprecipitations.
      2) Transfer the lysate to a 1.5 ml microcentrifuge tube and centrifuge at 15,000 X g for 10 min at 4°C. Save supernatant–this is the lysate.
      3) Pre-clear the 1.5 ml lysate with 10 µg normal mouse IgG (Cat. No. NI03) and 250 µl of Protein A sepharose solution (Cat. No. IP02). Incubate at 4°C for 60 min with continuous end-over-end mixing.
      4) Pellet the Protein A Sepharose/mouse IgG complexes by centrifugation at 1000 X g for 1 min. Save the supernatant–this is the pre-cleared lysate.
      5) Add 2 µg of APC (Ab-5) to 300 µl of the pre-cleared lysate and incubate at 4°C for 60 min with continuous end-over-end mixing.
      6) Add 50 µl of Protein A sepharose and incubate at 4°C for 60 min with continuous end-over-end mixing.
      7) Pellet the Protein A sepharose/immunocomplexes by centrifugation at 1000 X g for 1 min. Discard the supernatant.
      8) Wash the Protein A sepharose/immunocomplex pellet 3 times with 1 ml of wash buffer.
      9) Resuspend pellet in 50 µl of 2X Laemmli sample buffer and heat at 100°C for 10 min.
      10) Detect APC proteins by immunoblotting as described for APC (Ab-1), Cat. No. OP44, or APC (Ab-2), Cat. No. OP47.
      Biological Information
      Immunogena recombinant protein consisting of the N-terminal 226 amino acids of APC
      ImmunogenHuman
      CloneCF11
      HostMouse
      IsotypeIgG2b
      Species Reactivity
      • Human
      • Mouse
      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
      Catalogue Number GTIN
      OP62 0

      Documentation

      Anti-APC (Ab-5) Mouse mAb (CF11) SDS

      Title

      Safety Data Sheet (SDS) 

      Anti-APC (Ab-5) Mouse mAb (CF11) Certificates of Analysis

      TitleLot Number
      OP62

      References

      Reference overview
      Smith, K.J., et al. 1993. Proc. Natl. Acad. Sci. USA 90, 2846.
      Su, L.-K., et al. 1993. Can. Res. 53, 2728.
      Boynton, R.F., et al. 1992. Proc. Natl. Acad. Sci. USA 89, 3385.
      D'Amico, D., et al. 1992. Cancer Res. 52, 1996.
      Fearon, E.R., and Jones, P.A. 1992. FASEB J. 6, 2783.
      Miyoshi, Y., et al. 1992. Proc. Natl. Acad. Sci. USA 89, 4452.
      Powell, S.M., et al. 1992. Nature 359, 235.
      Groden, J., et al. 1991. Cell 66, 589.
      Kinzler, K.W., et al. 1991. Science 253, 661.
      Nishisho, I., et al. 1991. Science 253, 665.
      Data Sheet

      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.

      Revision10-May-2010 RFH
      SynonymsAnti-Adenomatous Polyposis Coli
      ApplicationImmunoprecipitation (see comments and application references)
      DescriptionPurified mouse monoclonal antibody generated by immunizing mice with the specified immunogen and fusing splenocytes with SP2/0 mouse myeloma cells. Recognizes various full-length and truncated forms of APC.
      BackgroundAnti-APC (Ab-5) is an IgG2bκ mouse monoclonal antibody raised against an epitope in the amino-terminal 226 amino acids of the 2843 amino acid APC protein p300APC. Colon cancer generally occurs sporadically although there is clear genetic predisposition in some families. Familial Adenomatous Polyposis Coli (FAP) is an autosomal dominantly inherited disorder that exhibits 80 to 100% penetrance and predisposes to colon cancer. The gene responsible for FAP has been assigned to chromosome 5q21, a region that is often lost in tumors from patients with sporadic colorectal cancer. Recently, a gene called APC, for adenomatous polyposis coli, from the 5q21 locus was isolated and shown to contain mutations in DNA prepared from tumors of patients with sporadic colorectal cancer and in germ line DNA and tumor DNA from FAP patients. In 79 unrelated patients with FAP, mutations in APC were found in 67% of the cases; and 49 of the 53 mutations were either small deletions/insertions or were point mutations that introduced stop codons into the reading frame. Furthermore, APC mutations have been found in the earliest tumors that could be analyzed (0.5 cm diameter) and the mutation frequency remained constant as tumors progressed from benign to malignant stages. Analysis of the APC protein demonstrates that inactivating mutations result in stable truncated proteins that can be identified in colon tumors from both FAP and sporadic colon cancer patients as well as in constitutional cells of FAP patients. Loss of heterozygosity at the APC locus has also been reported for esophageal cancer and lung cancer.
      HostMouse
      Immunogen speciesHuman
      Immunogena recombinant protein consisting of the N-terminal 226 amino acids of APC
      CloneCF11
      IsotypeIgG2b
      Specieshuman, mouse
      Positive controlHEK293, A-431, HCT116, or HT-1080 cells (full-length APC) or SW480 cells (truncated APC)
      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
      CommentsUsing this antibody various truncated and full lenghth forms of the APC protein have been observed in tumor cell lines and in lymphocytes obtained from FAP patients by immunoprecipiation. APC expression and turnover are very low making metabolic labelling difficult. APC immunocomplexes can be immunoprecipitated using Anti-APC (Ab-5) and detected in a immunoblot using Anti-APC (Ab-1), Cat. No. OP44 or Anti-APC (Ab-2), Cat. No. OP47L.

      IMMUNOPRECIPITATION PROCEDURE

      Can be used to immunoprecipitate full length and truncated APC protein APC. Precipitated APC proteins should then be visualized using APC (Ab-1) or APC (Ab-2) and the procedure provided with those products. APC expression and turnover are very low, therefore, immunoprecipitation of metabolically labeled lysates is not recommended.

      Materials:

      Equipment
      • Rocking Platform

      Solutions and Reagents:
      • Anti-APC (Ab-5), Mouse mAb (CF11) Cat. No. OP62
      • Protein A Sepharose swollen 1:1 in Wash Buffer
      • Normal Mouse IgG
      • Lysis Buffer: 100 mM NaCl, 50 mM Tris-HCl pH 7.5, 0.5% NP-40 Alternative, 0.5 µg/ml Leupeptin, 1.0 µg/ml Pepstatin, 0.2 mM PMSF
      • Wash Buffer: 100 mM NaCl, 50 mM Tris-HCl pH 7.5, 0.5% NP-40 Alternative
      • 2X Laemmli Sample Buffer: 120 mM Tris-HCl pH 6.8, 4% SDS, 20% glycerol, 200 mM DTT, 0.01% Bromophenol Blue

      Procedure:
      1) Lyse cells in lysis buffer. For adherent cells, lyse an 80-90% confluent 10 cm plate in 1.5 ml of buffer. Incubate at 4°C for 30 min with rocking. This will make enough lysate for 5 immunoprecipitations.
      2) Transfer the lysate to a 1.5 ml microcentrifuge tube and centrifuge at 15,000 X g for 10 min at 4°C. Save supernatant–this is the lysate.
      3) Pre-clear the 1.5 ml lysate with 10 µg normal mouse IgG (Cat. No. NI03) and 250 µl of Protein A sepharose solution (Cat. No. IP02). Incubate at 4°C for 60 min with continuous end-over-end mixing.
      4) Pellet the Protein A Sepharose/mouse IgG complexes by centrifugation at 1000 X g for 1 min. Save the supernatant–this is the pre-cleared lysate.
      5) Add 2 µg of APC (Ab-5) to 300 µl of the pre-cleared lysate and incubate at 4°C for 60 min with continuous end-over-end mixing.
      6) Add 50 µl of Protein A sepharose and incubate at 4°C for 60 min with continuous end-over-end mixing.
      7) Pellet the Protein A sepharose/immunocomplexes by centrifugation at 1000 X g for 1 min. Discard the supernatant.
      8) Wash the Protein A sepharose/immunocomplex pellet 3 times with 1 ml of wash buffer.
      9) Resuspend pellet in 50 µl of 2X Laemmli sample buffer and heat at 100°C for 10 min.
      10) Detect APC proteins by immunoblotting as described for APC (Ab-1), Cat. No. OP44, or APC (Ab-2), Cat. No. OP47.
      Storage +2°C to +8°C
      Do Not Freeze Yes
      Toxicity Standard Handling
      ReferencesSmith, K.J., et al. 1993. Proc. Natl. Acad. Sci. USA 90, 2846.
      Su, L.-K., et al. 1993. Can. Res. 53, 2728.
      Boynton, R.F., et al. 1992. Proc. Natl. Acad. Sci. USA 89, 3385.
      D'Amico, D., et al. 1992. Cancer Res. 52, 1996.
      Fearon, E.R., and Jones, P.A. 1992. FASEB J. 6, 2783.
      Miyoshi, Y., et al. 1992. Proc. Natl. Acad. Sci. USA 89, 4452.
      Powell, S.M., et al. 1992. Nature 359, 235.
      Groden, J., et al. 1991. Cell 66, 589.
      Kinzler, K.W., et al. 1991. Science 253, 661.
      Nishisho, I., et al. 1991. Science 253, 665.
      Application referencesImmunoprecipitation Su, L.-K., et al. 1993 Can. Res. 53, 2728.