重组Anti-Eg5抗体[EPR23276-52] (ab254298)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23276-52] to Eg5
- Suitable for: Flow Cyt (Intra), ICC/IF, IP, WB, IHC-P
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-Eg5抗体[EPR23276-52]
参阅全部 Eg5 一抗 -
描述
兔单克隆抗体[EPR23276-52] to Eg5 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), ICC/IF, IP, WB, IHC-Pmore details
不适用于: IHC-Fr -
种属反应性
与反应: Mouse, 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: Human testis and tonsil tissue lysate. Mouse testis tissue lysate. Jurkat, NCI-H1975, MCF7 and 4T1 whole cell lysate. IHC-P: Human tonsil and testis tissue. Human breast cancer tissue. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt (intra): HeLa and NIH/3T3 cells. IP: HeLa and NIH/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: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR23276-52 -
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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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab254298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 |
---|---|---|
Flow Cyt (Intra) |
1/600.
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|
ICC/IF |
1/50.
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|
IP |
1/30.
|
|
WB |
1/1000. Predicted molecular weight: 119 kDa.
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|
IHC-P |
1/4000. 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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Flow Cyt (Intra)
1/600. |
ICC/IF
1/50. |
IP
1/30. |
WB
1/1000. Predicted molecular weight: 119 kDa. |
IHC-P
1/4000. 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
Motor protein required for establishing a bipolar spindle. Blocking of KIF11 prevents centrosome migration and arrest cells in mitosis with monoastral microtubule arrays. -
疾病相关
Defects in KIF11 are the cause of microcephaly with or without chorioretinopathy, lymphedema, or mental retardation (MCLMR) [MIM:152950]. An autosomal dominant disorder that involves an overlapping but variable spectrum of central nervous system and ocular developmental anomalies. Microcephaly ranges from mild to severe and is often associated with mild to moderate developmental delay and a characteristic facial phenotype with upslanting palpebral fissures, broad nose with rounded tip, long philtrum with thin upper lip, prominent chin, and prominent ears. Chorioretinopathy is the most common eye abnormality, but retinal folds, microphthalmia, and myopic and hypermetropic astigmatism have also been reported, and some individuals have no overt ocular phenotype. Congenital lymphedema, when present, is typically confined to the dorsa of the feet, and lymphoscintigraphy reveals the absence of radioactive isotope uptake from the webspaces between the toes. -
序列相似性
Belongs to the kinesin-like protein family. BimC subfamily.
Contains 1 kinesin-motor domain. -
翻译后修饰
Phosphorylated exclusively on serine during S phase, but on both serine and Thr-926 during mitosis, so controlling the association of KIF11 with the spindle apparatus (probably during early prophase). Phosphorylated upon DNA damage, probably by ATM or ATR.
A subset of this protein primarily localized at the spindle pole is phosphorylated by NEK6 during mitosis; phosphorylation is required for mitotic function. -
细胞定位
Cytoplasm. Cytoplasm > cytoskeleton > spindle pole. - Information by UniProt
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数据库链接
- Entrez Gene: 3832 Human
- Entrez Gene: 16551 Mouse
- Omim: 148760 Human
- SwissProt: P52732 Human
- SwissProt: Q6P9P6 Mouse
- Unigene: 8878 Human
- Unigene: 42203 Mouse
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别名
- EG5 antibody
- HKSP antibody
- KIF11 antibody
see all
图片
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Immunohistochemical analysis of paraffin-embedded human testis tissue labeling Eg5 with 254298 at 1/1/4000 (0.17µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in spermatocytes of human testis is observed.
The section was incubated with ab254298 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with hematoxylin.Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 cells labeling Eg5 with ab254298 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 µg/ml dilution (Green). Confocal image showing cytoplasmic and spindle (arrow) staining in NIH/3T3 cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 µg/ml dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 2 µg/ml dilution.
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All lanes : Anti-Eg5 antibody [EPR23276-52] (ab254298) at 1/1000 dilution
Lane 1 : Human testis tissue lysate
Lane 2 : Human tonsil tissue lysate
Lane 3 : Human lung tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/100000 dilution
Predicted band size: 119 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile observed is consistent with what has been described in the literature (PMID:23857769; 27492783).
Negative control: Human lung (PMID: 27279560).
The minor bands beneath the target band may correspond to degradation.
Exposure time: 48 seconds.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labeling Eg5 with ab254298 at 1/600 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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Eg5 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast), whole cell lysate 10 ug with ab254298 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254298 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: NIH/3T3 whole cell lysate 10 ug.
Lane 2: ab254298 IP in NIH/3T3 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab254298 in NIH/3T3 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes
Fresh lysates were used in this IP.
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Eg5 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate 10 ug with ab254298 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254298 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: HeLa whole cell lysate 10 ug.
Lane 2: ab254298 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab254298 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
Fresh lysates were used in this IP.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Eg5 with ab254298 at 1/600 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling Eg5 with ab254298 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 µg/ml dilution (Green). Confocal image showing cytoplasmic and spindle (arrow) staining in HeLa cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 µg/ml dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 2 µg/ml dilution.
-
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling Eg5 with ab254298 at 1/4000 (0.17µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Scattered cytoplasmic staining in human breast cancer cells is observed (PMID:29181100).
The section was incubated with ab254298 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with hematoxylin.Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Eg5 with 254298 at 1/4000 (0.17µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in human tonsil is observed (PMID:25277178).
The section was incubated with ab254298 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with hematoxylin.Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
All lanes : Anti-Eg5 antibody [EPR23276-52] (ab254298) at 1/1000 dilution
Lane 1 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
Lane 2 : NCI-H1975 (human adenocarcinoma lung epithelial cell)
Lane 3 : MCF7 (human breast adenocarcinoma epithelial cell)
Lane 4 : 4T1 (mouse mammary gland carcinoma epithelial cell)
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: 119 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile observed is consistent with what has been described in the literature (PMID:23857769; 27492783).
Fresh lysates were tested in this WB.
The minor bands beneath the target band in lane 1 and lane 4 may correspond to degradation.
Exposure time: 15 seconds.
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All lanes : Anti-Eg5 antibody [EPR23276-52] (ab254298) at 1/1000 dilution
Lane 1 : Mouse testis tissue lysate
Lane 2 : Mouse brain tissue 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: 119 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST.
Negative control: Mouse brain (PMID: 17974955).
The minor bands beneath the target band may correspond to degradation.
Exposure time: 48 seconds.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (2)
ab254298 被引用在 2 文献中.
- Yu WX et al. Kinesin-5 Eg5 is essential for spindle assembly, chromosome stability and organogenesis in development. Cell Death Discov 8:490 (2022). PubMed: 36513626
- Liu G et al. CDK6 is stimulated by hyperthermia and protects gastric cancer cells from hyperthermia-induced damage. Mol Med Rep 23:N/A (2021). PubMed: 33537819