Anti-PLK1抗体[36-298] (ab17057)
Key features and details
- Mouse monoclonal [36-298] to PLK1
- Suitable for: Flow Cyt (Intra), ICC, WB
- Reacts with: Mouse, Human, Recombinant fragment
- Isotype: IgG1
概述
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产品名称
Anti-PLK1抗体[36-298]
参阅全部 PLK1 一抗 -
描述
小鼠单克隆抗体[36-298] to PLK1 -
宿主
Mouse -
经测试应用
适用于: Flow Cyt (Intra), ICC, WBmore details -
种属反应性
与反应: Mouse, Human, Recombinant fragment
预测可用于: Rat -
免疫原
Recombinant full length protein corresponding to Human PLK1. His-PLK1 full length purified from Sf9 cells.
Database link: P53350 -
表位
aa330-370. -
阳性对照
- WB: HEK-293, HeLa S3 or U-2 OS cell lysate ICC: HeLa, HeLa S3, NIH/3T3 or U-2 OS cells Flow Cyt (Intra): HeLa cells.
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常规说明
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 or -80°C. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
36-298 -
同种型
IgG1 -
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
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab17057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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Flow Cyt (Intra) |
Use 5µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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ICC |
Use a concentration of 1 µg/ml.
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WB | (1) |
Use a concentration of 1 µg/ml. Detects a band of approximately 66 kDa (predicted molecular weight: 68 kDa).
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说明 |
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Flow Cyt (Intra)
Use 5µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
ICC
Use a concentration of 1 µg/ml. |
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 66 kDa (predicted molecular weight: 68 kDa). |
靶标
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功能
Serine/threonine-protein kinase that performs several important functions throughout M phase of the cell cycle, including the regulation of centrosome maturation and spindle assembly, the removal of cohesins from chromosome arms, the inactivation of APC/C inhibitors, and the regulation of mitotic exit and cytokinesis. Required for recovery after DNA damage checkpoint and entry into mitosis. Required for kinetochore localization of BUB1B. Phosphorylates SGOL1. Required for spindle pole localization of isoform 3 of SGOL1 and plays a role in regulating its centriole cohesion function. Phosphorylates BORA, and thereby promotes the degradation of BORA. Contributes to the regulation of AURKA function. Regulates TP53 stability through phosphorylation of TOPORS. -
组织特异性
Placenta and colon. -
序列相似性
Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. CDC5/Polo subfamily.
Contains 2 POLO box domains.
Contains 1 protein kinase domain. -
发展阶段
Accumulates to a maximum during the G2 and M phases, declines to a nearly undetectable level following mitosis and throughout G1 phase, and then begins to accumulate again during S phase. -
翻译后修饰
Catalytic activity is enhanced by phosphorylation of Thr-210. Phosphorylation at Thr-210 is first detected on centrosomes in the G2 phase of the cell cycle, peaks in prometaphase and gradually disappears from centrosomes during anaphase.
Autophosphorylation and phosphorylation of Ser-137 may not be significant for the activation of PLK1 during mitosis, but may enhance catalytic activity during recovery after DNA damage checkpoint.
Ubiquitinated by the anaphase promoting complex/cyclosome (APC/C) in anaphase and following DNA damage, leading to its degradation by the proteasome. Ubiquitination is mediated via its interaction with FZR1/CDH1. Ubiquitination and subsequent degradation prevents entry into mitosis and is essential to maintain an efficient G2 DNA damage checkpoint. -
细胞定位
Nucleus. Chromosome > centromere > kinetochore. Cytoplasm > cytoskeleton > centrosome. During early stages of mitosis, the phosphorylated form is detected on centrosomes and kinetochores. Localizes to the outer kinetochore. Presence of SGOL1 and interaction with the phosphorylated form of BUB1 is required for the kinetochore localization. - Information by UniProt
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数据库链接
- Entrez Gene: 5347 Human
- Entrez Gene: 18817 Mouse
- Entrez Gene: 25515 Rat
- Omim: 602098 Human
- SwissProt: P53350 Human
- SwissProt: Q07832 Mouse
- SwissProt: Q62673 Rat
- Unigene: 592049 Human
see all -
别名
- Cell cycle regulated protein kinase antibody
- PLK 1 antibody
- PLK antibody
see all
图片
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ab17057 staining PLK1 in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab17057 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
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All lanes : Anti-PLK1 antibody [36-298] (ab17057)
Lane 1 : Recombinant PLK1
Lane 2 : U-2 OS (Human bone osteosarcoma epithelial cell line) cell extract
Lane 3 : HeLa S3 cell extract
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 66 kDa why is the actual band size different from the predicted?10% SDS-PAGE gel.
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Immunofluoresence using ab17057 and either HeLa S3, NIH/3T3 (Mouse embyro fibroblast cell line) or U-2 OS (Human bone osteosarcoma epithelial cell line) cells.
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Anti-PLK1 antibody [36-298] (ab17057) at 1 µg/ml + HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate at 20 µg
Secondary
Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 66 kDa why is the actual band size different from the predicted?
Exposure time: 2 minutes -
In panel one HeLa (Human epithelial cell line from cervix adenocarcinoma) cells were stained with ab17057 (green) and DAPI. In the second panel, cells were stained with ab17057 (green) and SH-CREST (red), which stains the centromeres. Fix 30 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with 1 mg/ml Na borohydride or 100 mM ammonium chloride in PEM. Permeabilized for 30 minutes with 0.5% TX-100 in PEM. Block 30 minutes in 5% milk in TBST. Primary antibody incubated overnight at 4°C diluted 1/400 in 5% milk in TBST. Secondary antibody incubated 1 hour at RT diluted in 5% milk in TBST. Post-fix 20 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with ammonium chloride in PEM. Counterstain with DAPI in TBST. Mount with ProLong Gold antifade reagent from Invitrogen.
Notes: Ample washing between each step.
TBST = Tris buffered saline + 0.1% Tween. PEM = 80 mM K-PIPES, pH 6.8, 5 mM EGTA, 2 mM MgCl2.
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Flow cytometry overlay histogram showing HeLa cells stained with ab17057 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab17057) (1x106 in 100 µl at 5 µg/ml) for 30 min at 22°C.
The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150117) was used at 1/4000 dilution for 30 min at 22°C.
Isotype control antibody (black line) was mouse IgG1 kappa; (ab170190) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (30)
ab17057 被引用在 30 文献中.
- Zhang Z et al. PLK1 Mitigates Intervertebral Disc Degeneration by Delaying Senescence of Nucleus Pulposus Cells. Front Cell Dev Biol 10:819262 (2022). PubMed: 35372354
- Ogonuki N et al. Birth of mice from meiotically arrested spermatocytes following biparental meiosis in halved oocytes. EMBO Rep 23:e54992 (2022). PubMed: 35587095
- Zeng TT et al. HN1L/AP-2γ/PLK1 signaling drives tumor progression and chemotherapy resistance in esophageal squamous cell carcinoma. Cell Death Dis 13:1026 (2022). PubMed: 36476988
- Tavernier N et al. Bora phosphorylation substitutes in trans for T-loop phosphorylation in Aurora A to promote mitotic entry. Nat Commun 12:1899 (2021). PubMed: 33771996
- Blengini CS et al. Aurora kinase A is essential for meiosis in mouse oocytes. PLoS Genet 17:e1009327 (2021). PubMed: 33901174