Anti-Coilin抗体[IH10] (ab87913)
Key features and details
- Mouse monoclonal [IH10] to Coilin
- Suitable for: Flow Cyt (Intra), ICC, IP, IHC-P, WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG2b
Related conjugates and formulations
概述
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产品名称
Anti-Coilin抗体[IH10]
参阅全部 Coilin 一抗 -
描述
小鼠单克隆抗体[IH10] to Coilin -
宿主
Mouse -
经测试应用
适用于: Flow Cyt (Intra), ICC, IP, IHC-P, WBmore details -
种属反应性
与反应: Human -
免疫原
Synthetic peptide corresponding to Human Coilin.
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阳性对照
- WB: HeLa, Jurkat, HepG2, Hek293, MCF7, Caco-2 and SHSY-5Y cell lysates. IHC-P: Human normal testis. ICC: HeLa 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: 6.97% L-Arginine, PBS -
Concentration information loading...
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纯度
IgG fraction -
克隆
单克隆 -
克隆编号
IH10 -
同种型
IgG2b -
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
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab87913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 1µg for 106 cells.
ab170192 - Mouse monoclonal IgG2b, 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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IP |
Use at an assay dependent concentration.
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IHC-P | (1) |
Use a concentration of 5 µg/ml.
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WB |
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 63 kDa).
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说明 |
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Flow Cyt (Intra)
Use 1µg for 106 cells. ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
ICC
Use a concentration of 1 µg/ml. |
IP
Use at an assay dependent concentration. |
IHC-P
Use a concentration of 5 µg/ml. |
WB
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 63 kDa). |
靶标
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功能
Is a component of the nuclear coiled bodies (CBS) which are involved in the function or assembly/disassembly of nucleoplasmic snRNPs. During mitosis, CBS disassemble, coinciding with a mitotic-specific phosphorylation of p80 coilin. -
组织特异性
Found in all the cell types examined. -
序列相似性
Belongs to the coilin family. -
细胞定位
Nucleus. Nuclear coiled body located in the interchromatin space between the nucleolus and the nucleus. - Information by UniProt
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数据库链接
- Entrez Gene: 8161 Human
- Omim: 600272 Human
- SwissProt: P38432 Human
- Unigene: 532795 Human
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别名
- CLN 80 antibody
- CLN80 antibody
- COIL antibody
see all
图片
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ab87913 staining Coilin in HeLa cells. The cells were fixed with 100% methanol (5 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 ab87913 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).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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All lanes : Anti-Coilin antibody [IH10] (ab87913) at 1/500 dilution
Lane 1 : Jurkat whole cell at 20 µg
Lane 2 : Wild-type HeLa cell lysate at 20 µg
Lane 3 : Empty
Lane 4 : COIL knockout HeLa cell lysate at 20 µg
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
Predicted band size: 63 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 12 minutesBlocking buffer: 3% Milk
Gel type: MOPS
The band observed in the CRISPR-Cas9 edited lysate lane below 75 kDa is likely to represent a truncated form of COIL. This has not been investigated further and the functional properties of the gene product have not been determined.
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ab87913(1/2000) staining Coilin in assynchronous HeLa cells (green). Cells were fixed in methanol (this image) or paraformaldehyde (see abreview image), permeabilized with 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.
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IHC image of Coilin staining in human testis formalin fixed paraffin embedded tissue section*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab87913, 3µg/ml overnight at +4°C. An HRP-conjugated secondary (ab97250, 1/500 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.
The inset negative control image is taken from an identical assay without primary antibody.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
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All lanes : Anti-Coilin antibody [IH10] (ab87913) at 5 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 5 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 6 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 7 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate
Lane 8 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 63 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Additional bands at: 28 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 20 minutes -
Overlay histogram showing HeLa cells stained with ab87913 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab87913, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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IHC image of Coilin staining in Human normal testis formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab87913, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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Coilin was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Mouse monoclonal to Coilin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab87913.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Band: 75kDa: Coilin
实验方案
数据表及文件
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Datasheet download
文献 (29)
ab87913 被引用在 29 文献中.
- Ulianov SV et al. Suppression of liquid-liquid phase separation by 1,6-hexanediol partially compromises the 3D genome organization in living cells. Nucleic Acids Res 49:10524-10541 (2021). PubMed: 33836078
- Cheng Y et al. N6-Methyladenosine on mRNA facilitates a phase-separated nuclear body that suppresses myeloid leukemic differentiation. Cancer Cell 39:958-972.e8 (2021). PubMed: 34048709
- Xu C et al. HnRNP F/H associate with hTERC and telomerase holoenzyme to modulate telomerase function and promote cell proliferation. Cell Death Differ 27:1998-2013 (2020). PubMed: 31863069
- Dopie J et al. Tyramide signal amplification mass spectrometry (TSA-MS) ratio identifies nuclear speckle proteins. J Cell Biol 219:N/A (2020). PubMed: 32609799
- Venit T et al. Transcriptional Profiling Reveals Ribosome Biogenesis, Microtubule Dynamics and Expression of Specific lncRNAs to be Part of a Common Response to Cell-Penetrating Peptides. Biomolecules 10:N/A (2020). PubMed: 33213097