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OP64 Anti-p21WAF1 (Ab-1) Mouse mAb (EA10)

This Anti-p21WAF1 (Ab-1) Mouse mAb (EA10) is validated for use in FC, Immunoblotting, IF, IP, Paraffin Sections for the detection of p21WAF1 (Ab-1).

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

Synonyms: Anti-CIP1, Anti-SD11, Anti-p21, Anti-WAF

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OP64
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Overview

Replacement Information

Key Spec Table

Species ReactivityHostAntibody Type
HMMonoclonal Antibody

Pricing & Availability

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OP64-100UG
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      OP64-20UG
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          		 Plastic ampoule 20 μg
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          Description
          OverviewRecognizes 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 NumberOP64
          Brand Family Calbiochem®
          SynonymsAnti-CIP1, Anti-SD11, Anti-p21, Anti-WAF
          Application Data
          Detection of p21 WAF1 by immunoblotting: Sample: Whole cell lysate (56 µg) from MCF-7 cells. Primary antibody: Anti-p21WAF1 (Ab-1) Mouse mAb (EA10) (Cat. No. OP64) (3 µg/ml). Secondary antibody: Anti-Mouse IgG (Goat) Peroxidase Conjugate. Detection: chemiluminescence.
          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.
          El-Deiry, W.S., et al. 1995. Cancer Res. 55, 2910.
          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, 0.2% gelatin, pH 7.5.
          Negative controlUnstimulated cells, unstimulated skin tissue, or SK-OV-3 cells
          Positive controlAny cell line expressing wild-type p53 (e.g. Hs27 or U2OS treated with DNA-damaging agents) or skin or colon tissue
          Preservative≤0.1% sodium azide
          Quality LevelMQ100
          Applications
          Application ReferencesEpitope Identification Patrick O'Connor (NCI, personal communication). Original Clone, Paraffin Sections, Frozen Sections El-Deiry, W.S., et al. 1995. Cancer Res. 55, 2910. Paraffin Sections, Antigen Retrieval Mouriaux, Frédéric, et al. 2000. Invest Ophthamol. Vis. Sci. 41, 2837. Flow Cytometry Carter, A., et al. 2004. Brit. J. Haematol. 127, 425.
          Key Applications Flow Cytometry
          Frozen Sections
          Immunoblotting (Western Blotting)
          Immunofluorescence
          Immunoprecipitation
          Paraffin Sections
          Application NotesFlow Cytometry (2 µg/ml or use Cat. No. OP64F; see application references)
          Frozen Sections (5 µg/ml or use Cat. No. OP64F)
          Immunoblotting (1-3 µg/ml)
          Immunofluorescence (1-5 µg/ml or use Cat. No. OP64F)
          Immunoprecipitation (2 µg/sample)
          Paraffin Sections (5 µg/ml or use OP64F; heat pre-treatment required; see comments and application references)
          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 WAF1 expression. Serum stimulation of quiescent cells will give low level WAF1 expression independent of p53 expression. Untreated cells will express little p21WAF1 and can 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. This antibody will immunoprecipitate p21WAF1 but not associated proteins. For immunoblotting applications, use a 0.22 µm filter and visualize by chemiluminescence. For staining paraffin sections, heating the tissue in 10 mM citrate buffer is required (see application references). 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. 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 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
          OP64-100UG 07790788053918
          OP64-20UG 04055977227291

          Documentation

          Anti-p21WAF1 (Ab-1) Mouse mAb (EA10) SDS

          Title

          Safety Data Sheet (SDS) 

          Anti-p21WAF1 (Ab-1) Mouse mAb (EA10) Certificates of Analysis

          TitleLot Number
          OP64

          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.
          El-Deiry, W.S., et al. 1995. Cancer Res. 55, 2910.
          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.

          Revision12-March-2008 JSW
          SynonymsAnti-CIP1, Anti-SD11, Anti-p21, Anti-WAF
          ApplicationFlow Cytometry (2 µg/ml or use Cat. No. OP64F; see application references)
          Frozen Sections (5 µg/ml or use Cat. No. OP64F)
          Immunoblotting (1-3 µg/ml)
          Immunofluorescence (1-5 µg/ml or use Cat. No. OP64F)
          Immunoprecipitation (2 µg/sample)
          Paraffin Sections (5 µg/ml or use OP64F; heat pre-treatment required; see comments and application references)
          Application Data
          Detection of p21 WAF1 by immunoblotting: Sample: Whole cell lysate (56 µg) from MCF-7 cells. Primary antibody: Anti-p21WAF1 (Ab-1) Mouse mAb (EA10) (Cat. No. OP64) (3 µg/ml). Secondary antibody: Anti-Mouse IgG (Goat) Peroxidase Conjugate. Detection: chemiluminescence.
          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 or U2OS treated with DNA-damaging agents) or skin or colon tissue
          Negative controlUnstimulated cells, unstimulated skin tissue, or SK-OV-3 cells
          FormLiquid
          FormulationIn 50 mM sodium phosphate buffer, 0.2% gelatin, pH 7.5.
          Concentration Label Please refer to vial label for lot-specific concentration
          Preservative≤0.1% sodium azide
          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 WAF1 expression. Serum stimulation of quiescent cells will give low level WAF1 expression independent of p53 expression. Untreated cells will express little p21WAF1 and can 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. This antibody will immunoprecipitate p21WAF1 but not associated proteins. For immunoblotting applications, use a 0.22 µm filter and visualize by chemiluminescence. For staining paraffin sections, heating the tissue in 10 mM citrate buffer is required (see application references). 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. Antibody should be titrated for optimal results in individual systems.
          Storage +2°C to +8°C
          Do Not Freeze Yes
          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.
          El-Deiry, W.S., et al. 1995. Cancer Res. 55, 2910.
          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 referencesEpitope Identification Patrick O'Connor (NCI, personal communication). Original Clone, Paraffin Sections, Frozen Sections El-Deiry, W.S., et al. 1995. Cancer Res. 55, 2910. Paraffin Sections, Antigen Retrieval Mouriaux, Frédéric, et al. 2000. Invest Ophthamol. Vis. Sci. 41, 2837. Flow Cytometry Carter, A., et al. 2004. Brit. J. Haematol. 127, 425.

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          Categories

          Life Science Research > Antibodies and Assays > Primary Antibodies