重组Anti-p21抗体[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
Related conjugates and formulations
概述
-
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
-
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
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.
性能
-
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
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
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. |
靶标
-
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. - 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
see all
图片
-
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 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
ab181602 was used as a GAPDH loading control.
-
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 secondsBlocking 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.
-
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.
-
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.
-
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.
-
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.
-
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 minutesBlocking and diluting buffer: 5% NDFM/TBST.
-
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 -
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) + 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.
数据表及文件
-
SDS download
-
Datasheet download
文献 (276)
ab109199 被引用在 276 文献中.
- Adili A et al. Atrial Fibrillation Underlies Cardiomyocyte Senescence and Contributes to Deleterious Atrial Remodeling during Disease Progression. Aging Dis 13:298-312 (2022). PubMed: 35111375
- Liu X et al. Characterization of transcriptional landscape in bone marrow-derived mesenchymal stromal cells treated with aspirin by RNA-seq. PeerJ 10:e12819 (2022). PubMed: 35127290
- Ma C et al. p21 restricts influenza A virus by perturbing the viral polymerase complex and upregulating type I interferon signaling. PLoS Pathog 18:e1010295 (2022). PubMed: 35180274
- Kim H et al. Attenuation of intrinsic ageing of the skin via elimination of senescent dermal fibroblasts with senolytic drugs. J Eur Acad Dermatol Venereol 36:1125-1135 (2022). PubMed: 35274377
- Dungan CM et al. Senolytic treatment rescues blunted muscle hypertrophy in old mice. Geroscience 44:1925-1940 (2022). PubMed: 35325353