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Knockout Tested Rabbit Recombinant Monoclonal p21 antibody. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 390 publications.


Images

Immunoprecipitation - Anti-p21 antibody [EPR362] (AB109520), expandable thumbnail
  • Western blot - Anti-p21 antibody [EPR362] (AB109520), expandable thumbnail
  • Western blot - Anti-p21 antibody [EPR362] (AB109520), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p21 antibody [EPR362] (AB109520), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-p21 antibody [EPR362] (AB109520), expandable thumbnail

Publications

  • Oncology letters 27:82023
    Effect of IFN‑γ encapsulated liposomes on major signal transduction pathways in the lymphocytes of patients with lung cancer.
    Applications:
    Unspecified application
    Reactive species:
    Unspecified reactive species
    Maysa Alhawamdeh,Belal Almajali,Wafa Hourani,Hamid Ali Nagi Al-Jamal,Abdullah Saleh Al-Wajeeh,Nesrin Riad Mwafi,Yousef Al-Hajaya,Hanan Kamel M Saad,Diana Anderson,Mahmoud Odeh,Ibraheam A Tarawneh
    PubMed 38028180
  • Aging 15:14607-146162023
    The protective effects of Phoenixin-20 in tumor necrosis factor α (TNF-α)-induced cell senescence of rheumatoid arthritis fibroblast-like synoviocytes (FLS).
    Applications:
    Unspecified application
    Reactive species:
    Unspecified reactive species
    Jinhua Yan,Ling Yao,Ying Tan,Yue Wang
    PubMed 38112587

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Tested

Tested
Tested

Species

Human

Dilution info

1/100 - 1/250

Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species

Human

Dilution info

1/10 - 1/100

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000 - 1/10000

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

For unpurified use at 1/50 - 1/100.

Tested
Tested

Species

Human

Dilution info

1/100

Notes

-

Associated Products

Select an associated product type

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      Target data

      Function

      May be involved in p53/TP53 mediated inhibition of cellular proliferation in response to DNA damage. Binds to and inhibits cyclin-dependent kinase activity, preventing phosphorylation of critical cyclin-dependent kinase substrates and blocking cell cycle progression. Functions in the nuclear localization and assembly of cyclin D-CDK4 complex and promotes its kinase activity towards RB1. At higher stoichiometric ratios, inhibits the kinase activity of the cyclin D-CDK4 complex. Inhibits DNA synthesis by DNA polymerase delta by competing with POLD3 for PCNA binding (PubMed:11595739). Plays an important role in controlling cell cycle progression and DNA damage-induced G2 arrest (PubMed:9106657).

      Alternative names

      Cyclin-dependent kinase inhibitor 1, CDK-interacting protein 1, Melanoma differentiation-associated protein 6, p21, MDA-6, CDKN1A, MDA6, CAP20, CDKN1, CIP1, PIC1, SDI1, WAF1

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      Knockout Tested Rabbit Recombinant Monoclonal p21 antibody. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 390 publications.

      Alternative names

      Cyclin-dependent kinase inhibitor 1, CDK-interacting protein 1, Melanoma differentiation-associated protein 6, p21, MDA-6, CDKN1A, MDA6, CAP20, CDKN1, CIP1, PIC1, SDI1, WAF1

      Key facts

      Isotype

      IgG

      Form

      Liquid

      Clonality

      Monoclonal

      Immunogen
      • The exact immunogen used to generate this antibody is proprietary information.
      Clone number

      EPR362

      Purification technique

      Affinity purification Protein A

      Specificity

      Expression levels of the target protein vary between different tissue/cell lines and in some cases induction may be required before a signal is observed.

      Concentration
      Loading...

      Storage

      Shipped at conditions

      Blue Ice

      Appropriate short-term storage duration

      1-2 weeks

      Appropriate short-term storage conditions

      +4°C

      Appropriate long-term storage conditions

      -20°C

      Aliquoting information

      Upon delivery aliquot

      Storage information

      Stable for 12 months at -20°C

      Notes

      Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

      This product is a recombinant monoclonal antibody, which offers several advantages including:

      • - High batch-to-batch consistency and reproducibility
      • - Improved sensitivity and specificity
      • - Long-term security of supply
      • - Animal-free batch production

      For more information, read more on recombinant antibodies.

      Supplementary info

      Activity summary

      The p21 protein also known as CDKN1A is an important regulator of cell cycle progression. It acts as a cyclin-dependent kinase inhibitor where it binds to and inhibits the activity of cyclin-CDK2 or -CDK4 complexes. The molecular weight of p21 is approximately 21 kDa. Cellular expression of p21 is diverse; it occurs in various tissues reflecting its broad role in maintaining cell cycle control. This protein's expression level is often assessed using techniques such as p21 western blotting p21 ELISA and p21 IHC staining.

      Biological function summary

      P21 plays a critical role in cell cycle regulation and DNA damage response. It associates with the p53 tumor suppressor protein forming an essential part of the p53 signaling pathway under stress conditions. p21 arrests cells in G1 phase allowing DNA repair or leading to cellular senescence. Through these mechanisms p21 integrates signals from stress and damage emphasizing its importance as a mediator in cellular checkpoint control.

      Pathways

      P21 is closely involved in the p53 and the Transforming Growth Factor-beta (TGF-β) signaling pathways. This integration helps in transmitting growth-inhibitory signals. p21 interacts with proteins such as p53 and cyclins ensuring precise regulation of the cell cycle and growth arrest when necessary. Triggered by p53 activation p21 contributes to preventing undisturbed cell proliferation acting as a safeguard against tumor formation.

      Associated diseases and disorders

      P21's deregulation associates with cancer and age-related diseases. Its impairment is a common feature in many cancers often linked with defects in the p53 pathway. Reduced p21 expression can contribute to unchecked cell division and tumor progression. Moreover in age-related diseases p21's role in promoting cellular senescence connects it with degenerative conditions. The intricate relationship between p21 and proteins such as p53 and cyclins highlights its potential as a therapeutic target for these disorders.

      Product promise

      We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

      In the unlikely event of one of our products not working as expected, you are covered by our product promise.

      Full details and terms and conditions can be found here:
      Terms & Conditions.

      15 product images

      • Immunoprecipitation - Anti-p21 antibody [EPR362] (ab109520), expandable thumbnail

        Immunoprecipitation - Anti-p21 antibody [EPR362] (ab109520)

        ab109520 (purified) at 1/50 immunoprecipitating p21 in HEK293 whole cell lysate.

        Lane 1 (input): HEK293 whole cell lysate (10μg)

        Lane 2 (+): ab109520 + HEK293 whole cell lysate (10μg).

        Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab109520 in HEK293 whole cell lysate.

        For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/1500 dilution.

        Blocking buffer and concentration: 5% NFDM/TBST.

        Diluting buffer and concentration: 5% NFDM /TBST.

        All lanes: Immunoprecipitation - Anti-p21 antibody [EPR362] (ab109520)

        Predicted band size: 18 kDa

        Observed band size: 21 kDa

      • Western blot - Anti-p21 antibody [EPR362] (ab109520), expandable thumbnail

        Western blot - Anti-p21 antibody [EPR362] (ab109520)

        False colour image of Western blot: Anti-p21 antibody [EPR362] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab109520 was shown to bind specifically to p21. A band was observed at 21 kDa in wild-type HeLa cell lysates with no signal observed at this size in CDKN1A knockout cell line Human CDKN1A knockout HeLa cell line ab255349 (knockout cell lysate ab263812). To generate this image, wild-type and CDKN1A knockout cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

        All lanes: Western blot - Anti-p21 antibody [EPR362] (ab109520) at 1/1000 dilution

        Lane 1: wild-type HeLa Vehicle Control Fluvastatin (20 uM, 24 h) cell lysate at 20 µg

        Lane 2: wild-type HeLa Treated Fluvastatin (50 uM, 24 h) cell lysate at 20 µg

        Lane 3: CDKN1A knockout HeLa Vehicle Control Fluvastatin (20 uM, 24 h) cell lysate at 20 µg

        Lane 4: CDKN1A knockout HeLa Treated Fluvastatin (50 uM, 24 h) cell lysate at 20 µg

        Lane 5: MCF7 cell lysate at 20 µg

        Lane 6: SH-SY5Y cell lysate at 20 µg

        Performed under reducing conditions.

        Predicted band size: 18 kDa

        Observed band size: 21 kDa

      • Western blot - Anti-p21 antibody [EPR362] (ab109520), expandable thumbnail

        Western blot - Anti-p21 antibody [EPR362] (ab109520)

        Lane 1: Wild-type DLD-1 cell lysate (20 μg)

        Lane 2: Wild-type DLD-1 20 μM 2,3-DCPE for 16hrs treated cell lysate (20 μg)

        Lane 3: p21 knockout DLD-1 cell lysate (20 μg)

        Lane 4: p21 knockout 20 μM 2,3-DCPE for 16hrs DLD-1 cell lysate (20 μg)

        Lane 5: HT1080 cell lysate (20 μg)


        Lanes 1 - 5: Merged signal (red and green). Green - ab109520 observed at 20 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

        ab109520 was shown to specifically recognize p21 in wild-type DLD-1 cells treated with 20 μM 2,3-DCPE. No band was observed when p21 knockout samples +/- 2,3-DCPE treatment were used. Wild-type and p21 knockout samples were subjected to SDS-PAGE. ab109520 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

        All lanes: Western blot - Anti-p21 antibody [EPR362] (ab109520)

        Predicted band size: 18 kDa

        Observed band size: 20 kDa

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p21 antibody [EPR362] (ab109520), expandable thumbnail

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p21 antibody [EPR362] (ab109520)

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid carcinoma tissue labelling p21 with purified ab109520 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

      • Immunocytochemistry/ Immunofluorescence - Anti-p21 antibody [EPR362] (ab109520), expandable thumbnail

        Immunocytochemistry/ Immunofluorescence - Anti-p21 antibody [EPR362] (ab109520)

        Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma cell line) labeling p21 with ab109520 at 1/500 (2 μg/ml). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, Alexa Fluor®488 Goat anti-Rabbit at 1/1000 (2 μg/ml) was used as secondary antibody. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 μg/ml) was used as counterstain AbID. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Nuclei were counter stained blue with DAPI.
        Confocal image showing nuclear staining on MCF7 cell line.

      • Flow Cytometry (Intracellular) - Anti-p21 antibody [EPR362] (ab109520), expandable thumbnail

        Flow Cytometry (Intracellular) - Anti-p21 antibody [EPR362] (ab109520)

        Overlay histogram showing HeLa cells stained with unpurified ab109520 (red line). The cells were fixed with 80% methanol (5 min) then permeabilized with 0.1% PBS-Tween 20 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab109520, 1/100) for 30 min at 22°C. The secondary antibody used was Alexa Fluorr® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

        Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

      • Western blot - Anti-p21 antibody [EPR362] (ab109520), expandable thumbnail

        Western blot - Anti-p21 antibody [EPR362] (ab109520)

        Different batches of ab109520 were tested on MCF7 (Human breast adenocarcinoma epithelial cell) lysate at 0.2 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 21 kDa.

        All lanes: Western blot - Anti-p21 antibody [EPR362] (ab109520)

        Predicted band size: 18 kDa

      • Immunocytochemistry/ Immunofluorescence - Anti-p21 antibody [EPR362] (ab109520), expandable thumbnail

        Immunocytochemistry/ Immunofluorescence - Anti-p21 antibody [EPR362] (ab109520)

        Immunocytochemistry/Immunofluorescence analysis of MCF7 cells labeling p21 with purified ab109520 at 1/1000. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.

        Control 1: primary antibody (1/1000) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

        Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

      • Western blot - Anti-p21 antibody [EPR362] (ab109520), expandable thumbnail

        Western blot - Anti-p21 antibody [EPR362] (ab109520)

        Lane 1: Wild-type DLD-1 cell lysate (20 μg)
        Lane 2: Wild-type DLD-1 20 μM 2,3-DCPE for 16hrs treated cell lysate (20 μg)
        Lane 3: p21 knockout DLD-1 cell lysate (20 μg)
        Lane 4: p21 knockout 20 μM 2,3-DCPE for 16hrs DLD-1 cell lysate (20 μg)
        Lane 5: HT1080 cell lysate (20 μg)

        Lanes 1 - 5: Merged signal (red and green). Green - ab109520 observed at 20 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

        This western blot image is a comparison between ab109520 and a competitor's top cited rabbit polyclonal antibody.

        All lanes: Western blot - Anti-p21 antibody [EPR362] (ab109520)

        Predicted band size: 18 kDa

      • Western blot - Anti-p21 antibody [EPR362] (ab109520), expandable thumbnail

        Western blot - Anti-p21 antibody [EPR362] (ab109520)

        Blocking and dilution buffer: 5% NFDM/TBST.

        All lanes: Western blot - Anti-p21 antibody [EPR362] (ab109520) at 1/2000 dilution

        Lane 1: MCF7 cell lysate at 20 µg

        Lane 2: HEK293 cell lysate at 20 µg

        Lane 3: U87-MG cell lysate at 20 µg

        Secondary

        All lanes: Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

        Predicted band size: 18 kDa, 24 kDa, 57 kDa

        Observed band size: 21 kDa

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p21 antibody [EPR362] (ab109520), expandable thumbnail

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p21 antibody [EPR362] (ab109520)

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human papillary carcinoma of the thyroid gland tissue labelling p21 with unpurified ab109520 at a dilution of 1/100.

        Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

      • Western blot - Anti-p21 antibody [EPR362] (ab109520), expandable thumbnail

        Western blot - Anti-p21 antibody [EPR362] (ab109520)

        Blocking and dilution buffer: 5% NFDM/TBST.

        All lanes: Western blot - Anti-p21 antibody [EPR362] (ab109520) at 1/10000 dilution

        All lanes: LnCaP cell lysate at 20 µg

        Secondary

        All lanes: Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

        Predicted band size: 18 kDa

        Observed band size: 21 kDa

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p21 antibody [EPR362] (ab109520), expandable thumbnail

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p21 antibody [EPR362] (ab109520)

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labelling p21 with unpurified ab109520 at a dilution of 1/100.

        Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

      • Western blot - Anti-p21 antibody [EPR362] (ab109520), expandable thumbnail

        Western blot - Anti-p21 antibody [EPR362] (ab109520)

        All lanes: Western blot - Anti-p21 antibody [EPR362] (ab109520) at 1/1000 dilution

        Lane 1: MCF7 cell lysate at 10 µg

        Lane 2: HeLa cell lysate at 10 µg

        Lane 3: HUVEC cell lysate at 10 µg

        Lane 4: LnCap cell lysate at 10 µg

        Lane 5: U87 MG cell lysate at 10 µg

        Lane 6: 293T cell lsyate at 10 µg

        Secondary

        All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721)

        Predicted band size: 18 kDa

      • Western blot - Anti-p21 antibody [EPR362] (ab109520), expandable thumbnail

        Western blot - Anti-p21 antibody [EPR362] (ab109520)

        Western blot: Anti-CDKN1A antibody [EPR362] (ab109520) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab109520 was shown to bind specifically to CDKN1A. A band was observed at 21 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CDKN1A knockout cell line. To generate this image, wild-type and CDKN1A knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

        All lanes: Western blot - Anti-p21 antibody [EPR362] (ab109520) at 1/1000 dilution

        Lane 1: Wild-type HCT 116 cell lysate at 20 µg

        Lane 2: CDKN1A knockout HCT 116 cell lysate at 20 µg

        Lane 3: Wild-type MCF7 cell lysate at 20 µg

        Lane 4: CDKN1A knockout MCF7 cell lysate at 20 µg

        Lane 5: Wild-type A549 cell lysate at 20 µg

        Lane 6: CDKN1A knockout A549 cell lysate at 20 µg

        Secondary

        All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

        Performed under reducing conditions.

        Predicted band size: 21 kDa

        Observed band size: 21 kDa

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