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OP64F Anti-p21WAF1 (58-77) (Ab-1) Mouse mAb (EA10) Fluorescein Conjugate

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OP64F
  
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      Overview

      Replacement Information

      Key Spec Table

      Species ReactivityHostAntibody Type
      HMMonoclonal Antibody
      Description
      Overview

      This product has been discontinued.



      Recognizes the ~21 kDa p21WAF1 protein in skin and colon tissue and in cells expressing wild-type p53 (e.g. Hs27 or U205 cells treated with DNA damaging agents).

      Catalogue NumberOP64F
      Brand Family Calbiochem®
      SynonymsAnti-SD11, Anti-p21, Anti-WAF, Anti-CIP1
      References
      ReferencesAgarwal, M.L., et al. 1995. Proc. Natl. Acad. Sci. USA 92, 8493.
      Chen, Y.Q., et al. 1995. Int. J. Oncology 7, 889.
      Deng, C., et al. 1995. Cell 82, 675.
      Waldman, T., et al. 1995. Cancer Res. 55, 5187.
      Elbendary, A., et al.1994. Cell Growth Diff. 5, 1301.
      El-Deiry, W.S., et al. 1994 Cancer Res. 54, 1169.
      Li, R., et al. 1994. Nature 371, 534.
      Michieli, P., et al. 1994. Cancer Res. 54, 3391.
      Noda, A., et al.1994. Exp. Cell Res. 211, 90.
      El-Deiry, W.S., et al.1993. Cell 75, 817.
      Gu, Y., et al. 1993. Nature 366, 707.
      Harper, J.W., et al.1993. Cell 75, 805.
      Xiong, Y., et al.1993. Genes Devel. 7, 1572.
      Xiong, Y., et al.1993. Nature 366, 701.
      Xiong, Y., et al.1992. Cell 71, 505.
      Product Information
      FormLiquid
      FormulationIn 50 mM sodium phosphate buffer, 50% glycerol.
      Negative controlUnstimulated cells, unstimulated skin tissue, or SK-OV-3 cells
      Positive controlAny cell line expressing wild-type p53 (e.g. Hs27 cells or U2OS cells treated with DNA damaging agents) or UV-treated skin or colon tissue
      PreservativeNone
      Quality LevelMQ100
      Applications
      Application ReferencesParaffin Sections, Frozen Sections, Original Clone El-Deiry, W.S., et al. 1995. Cancer Res. 55, 2910. Epitope Identification Patrick O'Connor (NCI, personal communication).
      Key Applications Flow Cytometry
      Immunofluorescence
      Application NotesFlow Cytometry (2 µg/ml)
      Frozen Sections (5 µg/ml)
      Immunoblotting (use Cat. No. OP64)
      Immunofluorescence (1-5 µg/ml)
      Immunoprecipitation (use Cat. No. OP64)
      Paraffin Sections (5 µg/ml, see comments)
      Application CommentsMaximal p21WAF1 expression requires wild type p53 activity. Treatment of U2OS or MCF7 cells with DNA damaging agents (such as doxorubicin at 0.2 µg/ml) induces wild type p53 expression which in turn activates p21WAF1 expression. Serum stimulation of quiescent cells will give low level p21WAF1 expression independent of p53 expression. Untreated cells will express little p21WAF1 and can also be used as a negative control. Cat. No. OP64F was tested in HALT cells induced by incubation at 31°C; FITC-goat anti-mouse IgG (Cat. No. DC13L) was used as a negative control. For staining paraffin sections, no pretreatment is usually needed, but some tissues may stain better after pretreating with heat or pressure cooker. In either paraffin or frozen sections of normal human colon, the non-dividing cells of colonic epithelium will stain positive for p21WAF1 while the proliferating compartment of crypts will not stain. For immunoprecipitation and immunoblotting use Cat. No. OP64. Antibody should be titrated for optimal results in individual systems.
      Biological Information
      Immunogenfull-length, recombinant, human p21WAF1
      ImmunogenHuman
      Epitopewithin amino acids 58-77 of human p21WAF1
      CloneEA10
      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 Dry Ice Only
      Toxicity Standard Handling
      Storage -20°C
      Protect from Light Protect from light
      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
      Catalogue Number GTIN
      OP64F 0

      Documentation

      Anti-p21WAF1 (58-77) (Ab-1) Mouse mAb (EA10) Fluorescein Conjugate SDS

      Title

      Safety Data Sheet (SDS) 

      Anti-p21WAF1 (58-77) (Ab-1) Mouse mAb (EA10) Fluorescein Conjugate Certificates of Analysis

      TitleLot Number
      OP64F

      References

      Reference overview
      Agarwal, M.L., et al. 1995. Proc. Natl. Acad. Sci. USA 92, 8493.
      Chen, Y.Q., et al. 1995. Int. J. Oncology 7, 889.
      Deng, C., et al. 1995. Cell 82, 675.
      Waldman, T., et al. 1995. Cancer Res. 55, 5187.
      Elbendary, A., et al.1994. Cell Growth Diff. 5, 1301.
      El-Deiry, W.S., et al. 1994 Cancer Res. 54, 1169.
      Li, R., et al. 1994. Nature 371, 534.
      Michieli, P., et al. 1994. Cancer Res. 54, 3391.
      Noda, A., et al.1994. Exp. Cell Res. 211, 90.
      El-Deiry, W.S., et al.1993. Cell 75, 817.
      Gu, Y., et al. 1993. Nature 366, 707.
      Harper, J.W., et al.1993. Cell 75, 805.
      Xiong, Y., et al.1993. Genes Devel. 7, 1572.
      Xiong, Y., et al.1993. Nature 366, 701.
      Xiong, Y., et al.1992. Cell 71, 505.
      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.

      Revision03-October-2007 RFH
      SynonymsAnti-SD11, Anti-p21, Anti-WAF, Anti-CIP1
      ApplicationFlow Cytometry (2 µg/ml)
      Frozen Sections (5 µg/ml)
      Immunoblotting (use Cat. No. OP64)
      Immunofluorescence (1-5 µg/ml)
      Immunoprecipitation (use Cat. No. OP64)
      Paraffin Sections (5 µg/ml, see comments)
      DescriptionPurified mouse monoclonal antibody generated by immunizing F1 mice with the specified immunogen and fusing splenocytes with SP2/0 mouse myeloma cells (see application references). Recognizes the ~21 kDa p21WAF1 protein.
      BackgroundThe tumor suppressor p53 transcriptionally activates a number of genes including the WAF1/CIP1 gene in response to DNA damage. The 21 kDa product of the WAF1 gene is found in a complex involving cyclins, cyclin dependent kinases (CDK), and PCNA in normal cells but not transformed cells and appears to be a universal inhibitor of CDK activity. One consequence of p21WAF1 binding to and inhibiting CDKs is to prevent CDK-dependent phosphorylation and subsequent inactivation of the Rb protein which is essential for cell cycle progression. p21WAF1 is, therefore, a potent and reversible inhibitor of cell cycle progression at both the G1 and G2 checkpoints, presumably to allow sufficient time for DNA repair to be completed. Irreversible G1 or G2 arrest leads to apoptosis. However, the role of p21WAF1 in apoptosis is less clear although p53-mediated apoptosis leads to increased WAF1 expression. Induction of p21WAF1 in response to DNA damage can occur by both p53-dependent and p53-independent mechanisms, in response to mitogenic stimuli, differentiation, or in tumor cells with mutated p53. Functional p21WAF1 is essential for p53-mediated G1 arrest presumably due to WAF1 inhibition of both CDK activity and PCNA-dependent DNA replication. WAF1 has also been identified as a gene involved in cellular senescence, termed sdi1. Not surprisingly, p21WAF1 overexpression is growth suppressive, consistent with its role as an inhibitor of CDKs. By inhibiting Rb inactivation in a p53-dependent fashion, p21WAF1 serves to integrate cell cycle control mediated by p53 and Rb.
      HostMouse
      Immunogen speciesHuman
      Immunogenfull-length, recombinant, human p21WAF1
      Epitopewithin amino acids 58-77 of human p21WAF1
      CloneEA10
      IsotypeIgG₁
      Specieshuman, not mouse, not rat
      Positive controlAny cell line expressing wild-type p53 (e.g. Hs27 cells or U2OS cells treated with DNA damaging agents) or UV-treated skin or colon tissue
      Negative controlUnstimulated cells, unstimulated skin tissue, or SK-OV-3 cells
      FormLiquid
      FormulationIn 50 mM sodium phosphate buffer, 50% glycerol.
      Concentration Label Please refer to vial label for lot-specific concentration
      PreservativeNone
      CommentsMaximal p21WAF1 expression requires wild type p53 activity. Treatment of U2OS or MCF7 cells with DNA damaging agents (such as doxorubicin at 0.2 µg/ml) induces wild type p53 expression which in turn activates p21WAF1 expression. Serum stimulation of quiescent cells will give low level p21WAF1 expression independent of p53 expression. Untreated cells will express little p21WAF1 and can also be used as a negative control. Cat. No. OP64F was tested in HALT cells induced by incubation at 31°C; FITC-goat anti-mouse IgG (Cat. No. DC13L) was used as a negative control. For staining paraffin sections, no pretreatment is usually needed, but some tissues may stain better after pretreating with heat or pressure cooker. In either paraffin or frozen sections of normal human colon, the non-dividing cells of colonic epithelium will stain positive for p21WAF1 while the proliferating compartment of crypts will not stain. For immunoprecipitation and immunoblotting use Cat. No. OP64. Antibody should be titrated for optimal results in individual systems.
      Storage Protect from light
      Avoid freeze/thaw
      -20°C
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-20°C).
      Toxicity Standard Handling
      ReferencesAgarwal, M.L., et al. 1995. Proc. Natl. Acad. Sci. USA 92, 8493.
      Chen, Y.Q., et al. 1995. Int. J. Oncology 7, 889.
      Deng, C., et al. 1995. Cell 82, 675.
      Waldman, T., et al. 1995. Cancer Res. 55, 5187.
      Elbendary, A., et al.1994. Cell Growth Diff. 5, 1301.
      El-Deiry, W.S., et al. 1994 Cancer Res. 54, 1169.
      Li, R., et al. 1994. Nature 371, 534.
      Michieli, P., et al. 1994. Cancer Res. 54, 3391.
      Noda, A., et al.1994. Exp. Cell Res. 211, 90.
      El-Deiry, W.S., et al.1993. Cell 75, 817.
      Gu, Y., et al. 1993. Nature 366, 707.
      Harper, J.W., et al.1993. Cell 75, 805.
      Xiong, Y., et al.1993. Genes Devel. 7, 1572.
      Xiong, Y., et al.1993. Nature 366, 701.
      Xiong, Y., et al.1992. Cell 71, 505.
      Application referencesParaffin Sections, Frozen Sections, Original Clone El-Deiry, W.S., et al. 1995. Cancer Res. 55, 2910. Epitope Identification Patrick O'Connor (NCI, personal communication).

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      Categories

      Life Science Research > Antibodies and Assays > Primary Antibodies