重组Anti-Geminin抗体[EPR14637] (ab195047)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR14637] to Geminin
- Suitable for: WB, ICC/IF, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-Geminin抗体[EPR14637]
参阅全部 Geminin 一抗 -
描述
兔单克隆抗体[EPR14637] to Geminin -
宿主
Rabbit -
经测试应用
适用于: WB, ICC/IF, IHC-Pmore details -
种属反应性
与反应: Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HepG2, HeLa (treated with thymidine), and HeLa whole cell lysate. IHC-P: Human triple-negative breast cancer and Human tonsil tissue. ICC/IF: MCF-7 cells
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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, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR14637 -
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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KO cell lines
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KO cell lysates
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab195047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 | (1) |
1/500. Predicted molecular weight: 24 kDa.
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ICC/IF | (2) |
1/100.
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IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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说明 |
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WB
1/500. Predicted molecular weight: 24 kDa. |
ICC/IF
1/100. |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
靶标
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功能
Inhibits DNA replication by preventing the incorporation of MCM complex into prereplication complex (pre-RC). It is degraded during the mitotic phase of the cell cycle. Its destruction at the metaphase-anaphase transition permits replication in the succeeding cell cycle. -
序列相似性
Belongs to the geminin family. -
发展阶段
Absent during G1 phase, accumulates during S, G2, and M phases, and disappears at the time of the metaphase-anaphase transition. - Information by UniProt
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数据库链接
- Entrez Gene: 51053 Human
- Omim: 602842 Human
- SwissProt: O75496 Human
- Unigene: 234896 Human
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别名
- DNA replication inhibitor antibody
- Gem antibody
- GEMI_HUMAN antibody
see all
图片
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All lanes : Anti-Geminin antibody [EPR14637] (ab195047) at 1/1000 dilution
Lane 1 : Untreated HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa treated with 2mM thymidine for 16 hours whole cell lysate
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: 24 kDa
Exposure time: 20 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
Geminin is regulated during apoptosis by site-specific cleavage, resulting in the generation of two lower molecular weight cleavage products (21,462 ±2.7 Da and 25,082 ±2 Da). (PMID: 17261582)
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All lanes : Anti-Geminin antibody [EPR14637] (ab195047) at 1/500 dilution
Lane 1 : Wild-type HeLa lysate
Lane 2 : Geminin DNA Replication Inhibitor knockout HeLa lysate
Lane 3 : HepG2 lysate
Lane 4 : U-87 MG lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 24 kDaLanes 1-4: Merged signal (red and green). Green - ab195047 observed at 32 kDa. Red - loading control ab7291 observed at 50 kDa.
ab195047 Anti-Geminin antibody [EPR14637] was shown to specifically react with Geminin DNA Replication Inhibitor in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264972 (knockout cell lysate ab257450) was used. Wild-type and Geminin DNA Replication Inhibitor knockout samples were subjected to SDS-PAGE. ab195047 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 500 and 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.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF-7 (Human breast adenocarcinoma cell line) cells labeling Geminin with ab195047 at 1/100 followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500(green). Confocal image showing nuclear staining on MCF-7 cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 (red).
The negative controls are as follows:-
-ve control 1 - ab195047 at 1/100 followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500.
-ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500. -
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Geminin with ab195047 at 1/500 followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Nucleus staining on Human tonsil tissue is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody and ab97051 at 1/500 as secondary antibody.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human triple-negative breast cancer tissue labeling Geminin with ab195047 at 1/500 followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Nucleus staining on Human triple-negative breast cancer tissue is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody and ab97051 at 1/500 as secondary antibody.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ab195047 staining Geminin in the HeLa cell line from Human cervix by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 Samples were incubated with primary antibody (1/50 in PBS ) for 1 hour at 22°C. Ab150081 (1/200) was used as the secondary antibody.
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 (7)
ab195047 被引用在 7 文献中.
- Vujovic F et al. The fate of notch-1 transcript is linked to cell cycle dynamics by activity of a natural antisense transcript. Nucleic Acids Res 49:10419-10430 (2021). PubMed: 34520549
- Baudoin NC et al. Asymmetric clustering of centrosomes defines the early evolution of tetraploid cells. Elife 9:N/A (2020). PubMed: 32347795
- Elbasani E et al. Kaposi's Sarcoma-Associated Herpesvirus Lytic Replication Is Independent of Anaphase-Promoting Complex Activity. J Virol 94:N/A (2020). PubMed: 32295923
- Chopra SS et al. Torin2 Exploits Replication and Checkpoint Vulnerabilities to Cause Death of PI3K-Activated Triple-Negative Breast Cancer Cells. Cell Syst 10:66-81.e11 (2020). PubMed: 31812693
- Su JH et al. Genome-Scale Imaging of the 3D Organization and Transcriptional Activity of Chromatin. Cell 182:1641-1659.e26 (2020). PubMed: 32822575
- Papp B et al. Genome-wide identification of direct RTA targets reveals key host factors for KSHV lytic reactivation. J Virol N/A:N/A (2018). PubMed: 30541837
- Pereira GC et al. Properties of LINE-1 proteins and repeat element expression in the context of amyotrophic lateral sclerosis. Mob DNA 9:35 (2018). PubMed: 30564290