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    coilin-antibody-ih10-ab87913.pdf

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Validated using a knockout cell line

Anti-Coilin antibody [IH10] (ab87913)

  • Datasheet
Reviews (3) Submit a question References (27)

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Immunocytochemistry - Anti-Coilin antibody [IH10] (ab87913)
  • Western blot - Anti-Coilin antibody [IH10] (ab87913)
  • Immunocytochemistry - Anti-Coilin antibody [IH10] (ab87913)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Coilin antibody [IH10] (ab87913)
  • Western blot - Anti-Coilin antibody [IH10] (ab87913)
  • Flow Cytometry - Anti-Coilin antibody [IH10] (ab87913)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Coilin antibody [IH10] (ab87913)
  • Immunoprecipitation - Anti-Coilin antibody [IH10] (ab87913)

Key features and details

  • Mouse monoclonal [IH10] to Coilin
  • Suitable for: ICC, IP, IHC-P, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG2b

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 HRP

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Human COIL (Coilin) knockout HeLa cell line (ab261757)

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Overview

  • Product name

    Anti-Coilin antibody [IH10]
    See all Coilin primary antibodies
  • Description

    Mouse monoclonal [IH10] to Coilin
  • Host species

    Mouse
  • Tested applications

    Suitable for: ICC, IP, IHC-P, Flow Cyt, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human Coilin.

  • Positive control

    • WB: HeLa, Jurkat, HepG2, Hek293, MCF7, Caco-2 and SHSY-5Y cell lysates. IHC-P: Human normal testis. ICC: HeLa cells. Flow Cyt: HeLa cells.
  • General notes

    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

Properties

  • Form

    Liquid
  • Storage instructions

    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.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: 6.97% L-Arginine, PBS
  • Concentration information loading...
  • Purity

    IgG fraction
  • Clonality

    Monoclonal
  • Clone number

    IH10
  • Isotype

    IgG2b
  • Research areas

    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Soma marker
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • RNA Processing
    • Other
    • Tags & Cell Markers
    • Subcellular Markers
    • Nucleus
    • Other Nuclear Bodies

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-Coilin antibody [IH10] (ab196714)
    • HRP Anti-Coilin antibody [IH10] (ab196715)
    • Alexa Fluor® 488 Anti-Coilin antibody [IH10] (ab197531)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

    • Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)
  • KO cell lines

    • Human COIL (Coilin) knockout HeLa cell line (ab261757)
  • KO cell lysates

    • Human COIL (Coilin) knockout HeLa cell lysate (ab257251)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab87913 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC
Use a concentration of 1 µg/ml.
IP
Use at an assay dependent concentration.
IHC-P (1)
Use a concentration of 5 µg/ml.
Flow Cyt
Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

WB
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 63 kDa).
Notes
ICC
Use a concentration of 1 µg/ml.
IP
Use at an assay dependent concentration.
IHC-P
Use a concentration of 5 µg/ml.
Flow Cyt
Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

WB
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 63 kDa).

Target

  • Function

    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.
  • Tissue specificity

    Found in all the cell types examined.
  • Sequence similarities

    Belongs to the coilin family.
  • Cellular localization

    Nucleus. Nuclear coiled body located in the interchromatin space between the nucleolus and the nucleus.
  • Target information above from: UniProt accession P38432 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 8161 Human
    • Omim: 600272 Human
    • SwissProt: P38432 Human
    • Unigene: 532795 Human
    • Alternative names

      • CLN 80 antibody
      • CLN80 antibody
      • COIL antibody
      • COIL_HUMAN antibody
      • Coilin antibody
      • Coilin p80 antibody
      • p80 antibody
      • P80 coilin antibody
      see all

    Images

    • Immunocytochemistry - Anti-Coilin antibody [IH10] (ab87913)
      Immunocytochemistry - Anti-Coilin antibody [IH10] (ab87913)

      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.

    • Western blot - Anti-Coilin antibody [IH10] (ab87913)
      Western blot - Anti-Coilin antibody [IH10] (ab87913)
      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 minutes


      Blocking 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.

    • Immunocytochemistry - Anti-Coilin antibody [IH10] (ab87913)
      Immunocytochemistry - Anti-Coilin antibody [IH10] (ab87913)This image is part of an abreview submitted by Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada

      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.

      See Abreview

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Coilin antibody [IH10] (ab87913)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Coilin antibody [IH10] (ab87913)

      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

    • Western blot - Anti-Coilin antibody [IH10] (ab87913)
      Western blot - Anti-Coilin antibody [IH10] (ab87913)
      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
    • Flow Cytometry - Anti-Coilin antibody [IH10] (ab87913)
      Flow Cytometry - Anti-Coilin antibody [IH10] (ab87913)
      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Coilin antibody [IH10] (ab87913)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Coilin antibody [IH10] (ab87913)

      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.

    • Immunoprecipitation - Anti-Coilin antibody [IH10] (ab87913)
      Immunoprecipitation - Anti-Coilin antibody [IH10] (ab87913)
      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

    Protocols

    • Flow cytometry protocols
    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (27)

    Publishing research using ab87913? Please let us know so that we can cite the reference in this datasheet.

    ab87913 has been referenced in 27 publications.

    • 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
    • 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
    • Ding H  et al. Systematic Analysis of Drug Vulnerabilities Conferred by Tumor Suppressor Loss. Cell Rep 27:3331-3344.e6 (2019). PubMed: 31189115
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-3 of 3 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Coilin antibody [IH10]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Bone Osteosarcoma Epithelial Cells)
    Permeabilization
    Yes - 0.3% tritonX-100
    Specification
    Bone Osteosarcoma Epithelial Cells
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 30 2021

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Coilin antibody [IH10]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Human brain motor cortex)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
    Specification
    Human brain motor cortex
    Blocking step
    Protein Block Novocastra from Leica Biosystems (# RE7102-CE) as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: RT°C
    Fixative
    Formalin
    Read More

    Mrs. Guinevere Lourenco

    Verified customer

    Submitted Nov 20 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Coilin antibody [IH10]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% Triton X100
    Read More

    DR. Kirk Mcmanus

    Verified customer

    Submitted Mar 11 2011

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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