Overview

  • Product name
    Anti-Nucleolin antibody [364-5]
    See all Nucleolin primary antibodies
  • Description
    Mouse monoclonal [364-5] to Nucleolin
  • Host species
    Mouse
  • Tested applications
    Suitable for: ICC/IF, IHC-P, WB, IP, Flow Cyt, In-Cell ELISAmore details
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat, Cow
  • Immunogen

    SU-DHL-1 Nuclei (Cell lysate nuclear fraction).

  • Positive control
    • HeLa and HDFn cells; HepG2, HDFn and SH-SY5Y whole cell lysates.
  • General notes

    This antibody clone is manufactured by Abcam.

    Product was previously marketed under the MitoSciences sub-brand.

     

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab136649 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/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 76 kDa.
IP Use a concentration of 1 µg/ml.
Flow Cyt Use a concentration of 1 µg/ml.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

In-Cell ELISA Use a concentration of 1 µg/ml.

Target

  • Function
    Nucleolin 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 similarities
    Contains 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 localization
    Nucleus > 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

Images

  • ab136649 staining Nucleolin in human breast cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 5% Triton X-100 in PBS and blocked with 5% BSA for 1 hour at 25°C. Samples were incubated with primary antibody (1/1000) for 1 hour. An undiluted Alexa Fluor® 546-conjugated anti-mouse IgG polyclonal was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-Nucleolin antibody [364-5] (ab136649) at 1 µg/ml

    Lane 1 : HepG2 whole cell lysate at 20 µg
    Lane 2 : HDFn whole cell lysate at 25 µg
    Lane 3 : SH-SY5Y whole cell lysate at 20 µg

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - AP at 1/3000 dilution

    Predicted band size: 76 kDa

  • Immunocytochemical analysis of HeLa cells (paraformaldehyde fixed (4%, 20 min) and Triton X-100 permeabilized (0.1%, 15 min)) labelling Nucleolin with ab136649 at 1µg/ml (red) and anti-GAPDH (ab110305) in green, as a counter stain.
    Secondary: Alexa 594 goat anti-mouse IgG1 at a 1/1000 dilution.
  • Immunocytochemical analysis of HDFn cells (paraformaldehyde fixed (4%, 20 min) and Triton X-100 permeabilized (0.1%, 15 min)) labelling Nucleolin with ab136649 at 1µg/ml (red) and anti-GAPDH (ab110305) in green, as a counter stain. Secondary: Alexa 594 goat anti-mouse IgG1 at a 1/1000 dilution.
  • Detection of Nucleolin by Immunoprecipitation.
    Nucleolin in HepG2 cell lysate (1 mg) was immunoprecipitated using 1 µg ab136649 antibody. The gel was stained with Coomassie brilliant blue G.
  • Flow Cytometric analysis of HeLa cells (fixed and permeabilized with methanol) labelling Nucleolin with ab136649 (blue) at 1µg/ml or an equal amount of an isotype control antibody (red).
  • IHC image of nucleolin staining in human liver HCC 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 ab136649, 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.

References

This product has been referenced in:
  • Son A  et al. PARN and TOE1 Constitute a 3' End Maturation Module for Nuclear Non-coding RNAs. Cell Rep 23:888-898 (2018). ICC/IF ; Human . Read more (PubMed: 29669292) »
  • Gao Z  et al. The PA-interacting host protein nucleolin acts as an antiviral factor during highly pathogenic H5N1 avian influenza virus infection. Arch Virol 163:2775-2786 (2018). Read more (PubMed: 29974255) »
See all 5 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Sample
Human Cell (Human Breast Cancer Line)
Specification
Human Breast Cancer Line
Permeabilization
Yes - 0.1% Triton x-100
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 13 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Sample
Human Cell (breast cell line)
Specification
breast cell line
Permeabilization
Yes - PBS 5%Triton- 5mn
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Mar 07 2014

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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