Name: Anti-p21 antibody [EPR3993]
Brand: RabMAb
SKU: ab109199
Rating: 4 (12 reviews)

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Western blot - Anti-p21 antibody [EPR3993] (ab109199)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR3993] to p21
Suitable for: WB
Knockout validated
Reacts with: Mouse, Rat, Human

Carrier Free

产品名称

Anti-p21抗体[EPR3993]
参阅全部 p21 一抗
描述

兔单克隆抗体[EPR3993] to p21
宿主

Rabbit
特异性

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 and primary cell samples.
We recommended ab109520 and ab188224 for use in IHC.
经测试应用

适用于: WBmore details
种属反应性

与反应: Mouse, Rat, Human
免疫原

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
阳性对照
- Raw 264.7, HCT116, MCF-7, PC-12 treated with 50ng/ml NFG for 48 hours whole cell lysate, wild-type HeLa Treated Fluvastatin (50 uM, 24 h) cell lysate, Wild-type DLD-1 20 µM 2,3-DCPE for 16hrs treated cell lysate
常规说明
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 production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

形式

Liquid
存放说明

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
存储溶液

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 59% PBS, 0.05% BSA
Concentration information loading...
纯度

Protein A purified
克隆

单克隆
克隆编号

EPR3993
同种型

IgG
研究领域

Alternative Versions
- Anti-p21 antibody [EPR3993] - BSA and Azide free (ab215971)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human CDKN1A knockout HCT116 cell line (ab266860)
KO cell lysates
- Human CDKN1A knockout HCT116 cell lysate (ab256870)
Positive Controls
- MCF7 whole cell lysate (ab29537)
Recombinant Protein
- Recombinant Human p21 protein (ab56278)
Related Products
- Prestained Protein Ladder - Broad molecular weight (10 - 245 kDa) (ab116028)

The Abpromise guarantee

Abpromise™承诺保证使用ab109199于以下的经测试应用

“应用说明”部分下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用	Ab评论	说明
WB	(6)	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.

说明
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.

功能

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.
组织特异性

Expressed in all adult human tissues, with 5-fold lower levels observed in the brain.
序列相似性

Belongs to the CDI family.
结构域

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.
翻译后修饰

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.
细胞定位

Cytoplasm. Nucleus.
Target information above from: UniProt accession P38936 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
数据库链接
- Entrez Gene: 1026 Human
- Entrez Gene: 12575 Mouse
- Entrez Gene: 114851 Rat
- Omim: 116899 Human
- SwissProt: P38936 Human
- SwissProt: P39689 Mouse
- Unigene: 370771 Human
- Unigene: 195663 Mouse
别名
- 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

Western blot - Anti-p21 antibody [EPR3993] (ab109199)

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

Lane 1 : Raw 264.7(Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 3 : C6 (Rat glial tumor glial cell) whole cell lysate
Lane 4 : PC-12(Rat adrenal gland pheochromocytoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 18 kDa
Observed band size: 18 kDa

Exposure time: 180 seconds

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

ab181602 was used as a GAPDH loading control.
Western blot - Anti-p21 antibody [EPR3993] (ab109199)

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

Lane 1 : PC-12(Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 2 : PC-12(Rat adrenal gland pheochromocytoma) treated with 50ng/ml NFG for 48 hours whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Developed using the ECL technique.

Predicted band size: 18 kDa
Observed band size: 18 kDa

Exposure time: 180 seconds

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

ab181602 was used as a GAPDH loading control.

We recommend using higher or super higher sensitivity ECL substrate for detecting.

Increase lysate amount can also help to get stronger signal.
Western blot - Anti-p21 antibody [EPR3993] (ab109199)

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

Lane 1 : wild-type HeLa Vehicle Control Fluvastatin (0 uM, 24 h) cell lysate
Lane 2 : wild-type HeLa Treated Fluvastatin (50 uM, 24 h) cell lysate
Lane 3 : CDKN1A knockout HeLa Vehicle Control Fluvastatin (0 uM, 24 h) cell lysate
Lane 4 : CDKN1A knockout HeLa Treated Fluvastatin (50 uM, 24 h) cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

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

False colour image of Western blot: Anti-p21 antibody [EPR3993] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab109199 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 CDKN2A knockout cell line ab255349 (knockout cell lysate ab263812). To generate this image, wild-type and CDKN2A 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
Western blot - Anti-p21 antibody [EPR3993] (ab109199)

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.
Western blot - Anti-p21 antibody [EPR3993] (ab109199)

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

Lane 1 : Wild-type HCT116 cell lysate
Lane 2 : CDKN1A knockout HCT116 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

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

Lanes 1- 2: Merged signal (red and green). Green - ab109199 observed at 20 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab109199 was shown to react with p21 in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266860 (knockout cell lysate ab256870) was used. Wild-Type HCT116 and CDKN1A knockout HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109199 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-p21 antibody [EPR3993] (ab109199)

Different batches of ab109199 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.
Western blot - Anti-p21 antibody [EPR3993] (ab109199)

Lane 1 : Anti-p21 antibody [EPR3993] (ab109199) (0.7ug/ul)
Lane 2 : Anti-p21 antibody [EPR362] (ab109520) (0.8ug/ul)

All lanes : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysates

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 18 kDa

Exposure time: 3 minutes

Blocking and diluting buffer: 5% NDFM/TBST.
Western blot - Anti-p21 antibody [EPR3993] (ab109199)

Lane 1 : Anti-p21 antibody [EPR3993] (ab109199) (1.4ug/ul)
Lane 2 : Anti-p21 antibody [EPR18021] (ab188224) (1.0ug/ul)

Lane 1 : RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
Lane 2 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 18 kDa
Western blot - Anti-p21 antibody [EPR3993] (ab109199)

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.
Western blot - Anti-p21 antibody [EPR3993] (ab109199)

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.
Anti-p21 antibody [EPR3993] (ab109199)

Western blot protocols

Click here to view the general protocols

SDS download

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Datasheet download

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发表研究结果有使用 ab109199？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab109199 被引用在 305 文献中.

Liu D et al. RRP Regulates Autophagy through the AMPK Pathway to Alleviate the Effect of Cell Senescence on Atherosclerosis. Oxid Med Cell Longev 2023:9645789 (2023). PubMed: 36756298
Liu Z et al. CD73/NT5E-mediated ubiquitination of AURKA regulates alcohol-related liver fibrosis via modulating hepatic stellate cell senescence. Int J Biol Sci 19:950-966 (2023). PubMed: 36778123
Li M et al. Genistein mitigates senescence of bone marrow mesenchymal stem cells via ERRα-mediated mitochondrial biogenesis and mitophagy in ovariectomized rats. Redox Biol 61:102649 (2023). PubMed: 36871183
Pillai PP et al. Involvement of lncRNA TUG1 in HIV-1 Tat-Induced Astrocyte Senescence. Int J Mol Sci 24:N/A (2023). PubMed: 36901763
Park J et al. CO-Induced TTP Activation Alleviates Cellular Senescence and Age-Dependent Hepatic Steatosis via Downregulation of PAI-1. Aging Dis 14:484-501 (2023). PubMed: 37008056

View all Publications for this product

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1-6 of 6 Abreviews or Q&A

Application

Western blot

Sample

Mouse Tissue lysate - whole (Heart)

Gel Running Conditions

Reduced Denaturing (4-12%)

Loading amount

30 µg

Specification

Heart

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Aug 10 2020

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

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Jul 10 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

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Sep 01 2016

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

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 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

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Christian Marx

Verified customer

提交于 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

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Christian Marx

Verified customer

提交于 Dec 02 2013

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Key features and details

Related conjugates and formulations

概述

产品名称

描述

宿主

特异性

经测试应用

种属反应性

免疫原

阳性对照

常规说明

性能

形式

存放说明

存储溶液

纯度

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Positive Controls

Recombinant Protein

Related Products

应用

The Abpromise guarantee

靶标

功能

组织特异性

序列相似性

结构域

翻译后修饰

细胞定位

数据库链接

别名

图片

实验方案

数据表及文件

SDS download

Datasheet download

文献 (305)

客户评价及客户问答

Western blot abreview for Anti-p21 antibody [EPR3993]

Western blot abreview for Anti-p21 antibody [EPR3993]

Western blot abreview for Anti-p21 antibody [EPR3993]

Western blot abreview for Anti-p21 antibody [EPR3993]

Western blot abreview for Anti-p21 [EPR3993] antibody

Western blot abreview for Anti-p21 [EPR3993] antibody