Overview

  • Product nameAnti-Nucleolin antibody
    See all Nucleolin primary antibodies
  • Description
    Rabbit polyclonal to Nucleolin
  • Tested applicationsSuitable for: WB, IP, IHC-Fr, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Nucleolin.

    (Peptide available as ab25315.)

  • Positive control
    • HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate, Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate, A431 (Human epithelial carcinoma cell line) Whole Cell Lysate, MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate, Brain (Mouse) Tissue Lysate, Pancreas (Mouse) Tissue Lysate, PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab22758 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 76 kDa (predicted molecular weight: 76 kDa).
IP Use at an assay dependent concentration.
IHC-Fr 1/400.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 µg/ml.

Target

  • FunctionNucleolin is the major nucleolar protein of growing eukaryotic cells. It is found associated with intranucleolar chromatin and pre-ribosomal particles. It induces chromatin decondensation by binding to histone H1. It is thought to play a role in pre-rRNA transcription and ribosome assembly. May play a role in the process of transcriptional elongation. Binds RNA oligonucleotides with 5'-UUAGGG-3' repeats more tightly than the telomeric single-stranded DNA 5'-TTAGGG-3' repeats.
  • Sequence similaritiesContains 4 RRM (RNA recognition motif) domains.
  • Post-translational
    modifications
    Some glutamate residues are glycylated by TTLL8. This modification occurs exclusively on glutamate residues and results in a glycine chain on the gamma-carboxyl group.
  • Cellular localizationNucleus > nucleolus. Cytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs.
  • Information by UniProt
  • Database links
  • Alternative names
    • C23 antibody
    • FLJ45706 antibody
    • MS1116 antibody
    • NCL antibody
    • Nucl antibody
    • NUCL_HUMAN antibody
    • Nucleolin antibody
    • Protein C23 antibody
    see all

Anti-Nucleolin antibody images

  • ICC/IF image of ab22758 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab22758, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • IHC image of Nucleolin staining in human tonsil FFPE section, performed on a 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 ab22758, 1µg/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.
  • All lanes : Anti-Nucleolin antibody (ab22758) at 1 µg/ml

    Lane 1 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 2 : Brain (Mouse) Tissue Lysate
    Lane 3 : Pancreas (Mouse) Tissue Lysate
    Lane 4 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Predicted band size : 76 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 60 kDa. We are unsure as to the identity of these extra bands.
  • Nucleolin was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to Nucleolin 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, Jurkat 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 ab22758.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 100kDa: Nucleolin.
  • Lane 1 : Marker
    Lanes 2 - 5 : Anti-Nucleolin antibody (ab22758) at 1 µg/ml

    Lane 1 : As above
    Lane 2 : Jurkat whole cell lysate (ab7899) at 20 µg
    Lane 3 : A431 whole cell lysate (ab7909) at 20 µg
    Lane 4 : Jurkat whole cell lysate (ab7899) at 20 µg with Human Nucleolin peptide (ab25315) at 1 µg/ml
    Lane 5 : A431 whole cell lysate (ab7909) at 20 µg with Human Nucleolin peptide (ab25315) at 1 µg/ml

    Secondary
    Lanes 2 - 5 : Goat polyclonal to Rabbit IgG H&L (HRP) Pre-Adsorbed at 1/10000 dilution


    Performed under reducing conditions.

    Predicted band size : 76 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)
  • ab22758 staining nucleolin in HeLa cells treated with Triptolide from Tripterygium wilfordii (ab120720), by ICC/IF. Changes in nuclear localization of nucleolin (from nuclelous to whole nucleous) correlates with increased concentration of Triptolide from Tripterygium wilfordii, as described in literature.
    The cells were incubated at 37°C for 1h in media containing different concentrations of ab120720 (Triptolide from Tripterygium wilfordii ) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab22758 (1 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody.

References for Anti-Nucleolin antibody (ab22758)

This product has been referenced in:
  • Das SK  et al. Poly(ADP-ribose) polymers regulate DNA topoisomerase I (Top1) nuclear dynamics and camptothecin sensitivity in living cells. Nucleic Acids Res 44:8363-75 (2016). Read more (PubMed: 27466387) »
  • Grondin Y  et al. Genetic Polymorphisms Associated with Hearing Threshold Shift in Subjects during First Encounter with Occupational Impulse Noise. PLoS One 10:e0130827 (2015). Read more (PubMed: 26121033) »

See all 26 Publications for this product

Product Wall

Application Western blot
Sample Human Cell lysate - whole cell (U2OS cells)
Gel Running Conditions Reduced Denaturing (4-12% Gradient Gel)
Loading amount 50 µg
Specification U2OS cells
Blocking step Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
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Submitted Oct 05 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (SHSY-5Y)
Permeabilization No
Specification SHSY-5Y
Fixative Methanol
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Submitted Aug 21 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step Background buster as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Sample Human Cell (Ovarian cancer cell line)
Specification Ovarian cancer cell line
Permeabilization Yes - 0.25% tritonx X-100
Fixative Formaldehyde
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Submitted Jun 02 2014

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3%
Sample Fruit fly (Drosophila melanogaster) Cell (adult testis)
Specification adult testis
Permeabilization Yes - .1% TritonX1000
Fixative Paraformaldehyde
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Submitted Jul 10 2013

Vielen Dank für Ihren Anruf.

Leider handelt handelt es sich bei dem monoklonen Antikörper hierbei nicht um ein In-Haus Produkt, so dass wir das Hybridoma nicht weiterverkaufen können. Des weiteren darf ich Ihnen aus betrieblichen Gründen nic...

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Thank you again for letting us know about this problem and for your patience while our lab is working with this antibody.
My colleagues in the lab have re-tested the antibody in mouse lung tissue, and we have also found that the apical staining is...

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Thank you for contacting us and your interest in our products.

As you have seen from the datasheet of the Anti-Nucleolin antibody (ab22758) we observe a consistent band of ˜100 kDa when performing Western blotting or immunoprecipitaion. We ha...

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Thank you very much for your interest in ab22758.

Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are cover...

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Thank you for your patience while I have been in touch with my colleague in our lab.
I have attached the human tonsil QC staining results from the lot that you most recently tried, and we will shortly send the QC results from the first lot that yo...

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Thank you for your reply.
I have let the lab know that they should re-test the antibody with mouse lung samples, and I will let you know as soon as I have the data. I will also send on the previous IHC images once I have access to them.
Pleas...

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1-10 of 13 Abreviews or Q&A

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