遺伝子ノックアウト細胞株 検証済
リコンビナント
RabMAb

Anti-p21 抗体 [EPR3993] (ab109199)

製品の概要

  • 製品名
    Anti-p21 antibody [EPR3993]
    p21 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EPR3993] to p21
  • 特異性
    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. This antibody is not recommended for use in WB with tissue samples. We recommended ab109520 and ab188224 for use in IHC.
  • アプリケーション
    適用あり: WB, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Human, African green monkey
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human p21 aa 100 to the C-terminus (C terminal).

  • ポジティブ・コントロール
    • WB: HUVEC, U87-MG, MCF-7, LnCaP, RAW264.7, PC-12 and C6 cell lysates.
  • 特記事項

     

    A trial size is available to purchase for this antibody.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab109199 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
WB 1/1000. Detects a band of approximately 21 kDa (predicted molecular weight: 18 kDa).

For unpurified use at 1/1000 - 1/10000. Not recommended for use with tissue samples.

ICC/IF Use at an assay dependent concentration.

ターゲット情報

  • 機能
    May be the important intermediate by which p53/TP53 mediates its role as an inhibitor 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.
  • 組織特異性
    Expressed in all adult human tissues, with 5-fold lower levels observed in the brain.
  • 配列類似性
    Belongs to the CDI family.
  • ドメイン
    The PIP-box K+4 motif mediates both the interaction with PCNA and the recuitment of the DCX(DTL) complex: while the PIP-box interacts with PCNA, the presence of the K+4 submotif, recruits the DCX(DTL) complex, leading to its ubiquitination.
    The C-terminal is required for nuclear localization of the cyclin D-CDK4 complex.
  • 翻訳後修飾
    Phosphorylation of Thr-145 by Akt or of Ser-146 by PKC impairs binding to PCNA. Phosphorylation at Ser-114 by GSK3-beta enhances ubiquitination by the DCX(DTL) complex.
    Ubiquitinated by MKRN1; leading to polyubiquitination and 26S proteasome-dependent degradation. Ubiquitinated by the DCX(DTL) complex, also named CRL4(CDT2) complex, leading to its degradation during S phase or following UV irradiation. Ubiquitination by the DCX(DTL) complex is essential to control replication licensing and is PCNA-dependent: interacts with PCNA via its PIP-box, while the presence of the containing the 'K+4' motif in the PIP box, recruit the DCX(DTL) complex, leading to its degradation.
  • 細胞内局在
    Cytoplasm. Nucleus.
  • Information by UniProt
  • 参照データベース
  • 別名
    • CAP20 antibody
    • CDK-interacting protein 1 antibody
    • CDKI antibody
    • CDKN1 antibody
    • Cdkn1a antibody
    • CDN1A_HUMAN antibody
    • CIP1 antibody
    • Cyclin Dependent Kinase Inhibitor 1A antibody
    • Cyclin-dependent kinase inhibitor 1 antibody
    • Cyclin-dependent kinase inhibitor 1A (P21) antibody
    • Cyclin-dependent kinase inhibitor 1A (p21, Cip1) antibody
    • DNA Synthesis Inhibitor antibody
    • MDA-6 antibody
    • MDA6 antibody
    • Melanoma differentiation-associated protein 6 antibody
    • Melanoma differentiation-associated protein antibody
    • p21 antibody
    • P21 protein antibody
    • p21CIP1 antibody
    • p21Cip1/Waf1 antibody
    • p21WAF antibody
    • PIC1 antibody
    • SDI1 antibody
    • SLC12A9 antibody
    • WAF1 antibody
    • Wild type p53 activated fragment 1 (WAF1) antibody
    • Wild type p53 activated fragment 1 antibody
    • Wildtype p53-activated fragment 1 antibody
    see all

画像



  • Predicted band size : 18 kDa
    Observed band size : 20 kDa (why is the actual band size different from the predicted?)

    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 - ab109199 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab109199 was shown to recognize p21 in WT DLD-1 cells with 2,3-DCPE treatment along with additional cross-reactive bands. When p21 knockout DLD-1 cells +/- 2,3-DCPE treatment were used, no band was observed. Wild-type and p21 knockout samples were subjected to SDS-PAGE. ab109199 and 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 (ab216773) and 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.

  • Immunocytochemistry/ Immunofluorescence analysis of HeLa cells labeling p21 with ab109199 at 1/400 dilution. Cells were fixed in paraformaldehyde and permeabilized with 0.5% Triton X-100 in PBS. Staining with ab109199 at 1/400 was carried out for 1 hour at 22°C in PBS buffer. ab150081, a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody was used at 1/200 dilution. DAPI was used to counterstain.

    See Abreview



  • Predicted band size : 18 kDa

    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 - ab109199 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.

    This western blot image is a comparison between ab109119 and a competitor's top cited rabbit polyclonal antibody.

  • Anti-p21 antibody [EPR3993] (ab109199) at 1/1000 dilution (purified) + MCF-7 cell lysate at 20 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 18 kDa
    Observed band size : 21 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-p21 antibody [EPR3993] (ab109199) at 1/5000 dilution (purified)

    Lane 1 : HUVEC cell lysate
    Lane 2 : U87-MG cell lysate
    Lane 3 : LnCaP cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 18 kDa
    Observed band size : 21 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Anti-p21 antibody [EPR3993] (ab109199) at 1/5000 dilution (purified) + RAW264.7 cell lysate at 20 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 18 kDa
    Observed band size : 21 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Anti-p21 antibody [EPR3993] (ab109199) at 1/1000 dilution (purified) + PC-12 cell lysate at 10 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 18 kDa
    Observed band size : 21 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-p21 antibody [EPR3993] (ab109199) at 1/1000 dilution (unpurified)

    Lane 1 : HUVEC cell lysate
    Lane 2 : U87MG cell lysate
    Lane 3 : C6 cell lysate

    Lysates/proteins at 10 µg per lane.


    Predicted band size : 18 kDa

    Secondary antibody - anti-rabbit HRP (ab6721).

プロトコール

参考文献

This product has been referenced in:
  • Song X  et al. Autophagy deficient keratinocytes display increased DNA damage, senescence and aberrant lipid composition after oxidative stress in vitro and in vivo. Redox Biol 11:219-230 (2017). WB ; Human . Read more (PubMed: 28012437) »
  • Cheng XY  et al. AMP-activated protein kinase reduces inflammatory responses and cellular senescence in pulmonary emphysema. Oncotarget 8:22513-22523 (2017). WB ; Mouse . Read more (PubMed: 28186975) »

See all 22 Publications for this product

レビューと Q&A

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse Derived Breast Cancer Cell Line)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
30 µg
Specification
Mouse Derived Breast Cancer Cell Line
Blocking step
Odyssey Blocking BUffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C
Username

Abcam user community

Verified customer

投稿 Jul 10 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Username

Dr. Kirk Mcmanus

Verified customer

投稿 Jan 16 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (MCF cells)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
25 µg
Specification
MCF cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

投稿 Sep 01 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (peritonei tumor)
Permeabilization
Yes - 0.3% Triton X 100
Specification
peritonei tumor
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Feb 24 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (LS174t)
Permeabilization
No
Specification
LS174t
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Aug 19 2015

Application
Western blot
Loading amount
50000 cells
Gel Running Conditions
Reduced Denaturing (10% gel)
Sample
Human Cell lysate - whole cell (MCF-7)
Specification
MCF-7
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

投稿 Aug 04 2014

Application
Western blot
Loading amount
15 µg
Gel Running Conditions
Reduced Denaturing (13%)
Sample
Human Cell lysate - whole cell (HeLa and HCT116 cell lysates)
Specification
HeLa and HCT116 cell lysates
Treatment
DMSO control (Ctl) and 7,5 ´M CPT-11 (CPT) for 24 hrs
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Christian Marx

Verified customer

投稿 Dec 02 2013

Application
Western blot
Loading amount
15 µg
Gel Running Conditions
Reduced Denaturing (13%)
Sample
Mouse Cell lysate - whole cell (mouse embryonic fibroblast (MEF) cell lysates)
Specification
mouse embryonic fibroblast (MEF) cell lysates
Treatment
DMSO control (Ctl) and 7,5 ´M CPT-11 (CPT) for 24 hrs
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Christian Marx

Verified customer

投稿 Dec 02 2013

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

登録