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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 Specifications Table

Species ReactivityHostAntibody Type
HMMonoclonal Antibody

Products

Catalog NumberPackaging Qty/Pack
OP64-100UG Plastic ampoule 100 μg
OP64-20UG Plastic ampoule 20 μg
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
Catalog 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.