重组Anti-p21抗体[EPR18021] (ab188224)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18021] to p21
- Suitable for: WB, IHC-P, IP, ICC/IF, Flow Cyt (Intra)
- Reacts with: Mouse
Related conjugates and formulations
概述
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产品名称
Anti-p21抗体[EPR18021]
参阅全部 p21 一抗 -
描述
兔单克隆抗体[EPR18021] 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 samples.
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经测试应用
适用于: WB, IHC-P, IP, ICC/IF, Flow Cyt (Intra)more details -
种属反应性
与反应: Mouse -
免疫原
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: RAW 264.7 whole cell lysate; NIH/3T3, untreated and treated with 1µM staurosporine for 2hrs, whole cell lysates. IHC-P: Mouse testis and lung tissues, mouse lung cancer tissue. ICC/IF: RAW 264.7 and NIH/3T3 cells. Flow Cyt (intra): RAW 264.7 cells. IP: NIN/3T3 whole cell lysate.
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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.
性能
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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 long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR18021 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab188224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 |
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WB | (1) |
1/1000. Detects a band of approximately 18 kDa (predicted molecular weight: 18 kDa).
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IHC-P | (5) |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
1/30.
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ICC/IF | (2) |
1/500.
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Flow Cyt (Intra) |
1/50.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
说明 |
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WB
1/1000. Detects a band of approximately 18 kDa (predicted molecular weight: 18 kDa). |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
1/30. |
ICC/IF
1/500. |
Flow Cyt (Intra)
1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
靶标
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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
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数据库链接
- Entrez Gene: 12575 Mouse
- SwissProt: P39689 Mouse
- Unigene: 195663 Mouse
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别名
- CAP20 antibody
- CDK-interacting protein 1 antibody
- CDKI antibody
see all
图片
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Immunohistochemical analysis of paraffin-embedded Mouse lung cancer tissue labeling p21 with ab188224 at 1/1000 dilution (0.517 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse lung cancer. The section was incubated with ab188224 for 30 mins at room temperature. The section was counterstained with Hematoxylin.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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All lanes : Anti-p21 antibody [EPR18021] (ab188224) at 1/1000 dilution
Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 1µM staurosporine for 2hrs, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 18 kDa
Observed band size: 18 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The expression level of p21 protein can be induced using staurosporine (protein kinase C inhibitor) (PMID:7677742).
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Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling p21 with ab188224 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear staining on mouse testis is observed (PMID: 9170103).
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling p21 with ab188224 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on RAW264.7 cells.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling p21with ab188224 at 1/50 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
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Anti-p21 antibody [EPR18021] (ab188224) at 1/1000 dilution + RAW264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 18 kDa
Observed band size: 18 kDa
Exposure time: 8 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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Different batches of ab188224 were tested on RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) lysate at 0.1 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 18 kDa.
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Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling p21 with ab188224 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Sporadic nuclear staining on mouse lung is observed (PMID: 25333671).
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling p21 with ab188224 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on NIH/3T3 cells.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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p21 was immunoprecipitated from 0.35 mg of NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate with ab188224 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab188224 at 1/500 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: NIH/3T3 whole cell lysate 10 µg (Input).
Lane 2: ab188224 IP in NIH/3T3 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab188224 in NIH/3T3 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (119)
ab188224 被引用在 119 文献中.
- Schloesser D et al. Senescent cells suppress macrophage-mediated corpse removal via upregulation of the CD47-QPCT/L axis. J Cell Biol 222:N/A (2023). PubMed: 36459066
- Chen M et al. Metformin protects lens epithelial cells against senescence in a naturally aged mouse model. Cell Death Discov 8:8 (2022). PubMed: 35013152
- Niwa Y et al. Cyclin D1 Binding Protein 1 Responds to DNA Damage through the ATM-CHK2 Pathway. J Clin Med 11:N/A (2022). PubMed: 35160302
- 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
- Marquez-Exposito L et al. Oxidative Stress and Cellular Senescence Are Involved in the Aging Kidney. Antioxidants (Basel) 11:N/A (2022). PubMed: 35204184