Key features and details
- Mouse monoclonal [364-5] to Nucleolin
- Suitable for: ICC/IF, IHC-P, WB, IP, Flow Cyt
- Reacts with: Human
- Isotype: IgG1
Product nameAnti-Nucleolin antibody [364-5]
See all Nucleolin primary antibodies
DescriptionMouse monoclonal [364-5] to Nucleolin
Tested applicationsSuitable for: ICC/IF, IHC-P, WB, IP, Flow Cytmore details
Species reactivityReacts with: Human
Does not react with: Mouse, Rat, Cow
Tissue, cells or virus. This information is considered to be commercially sensitive.
- HeLa and HDFn cells; HepG2, HDFn and SH-SY5Y whole cell lysates.
This antibody clone is manufactured by Abcam.
Product was previously marketed under the MitoSciences sub-brand.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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, as well as customer reviews and Q&As.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.30
Preservative: 0.02% Sodium azide
Constituents: 0.88% Sodium chloride, 0.36% HEPES
Concentration information loading...
Purification notesab136649 is purified by biochemical fractionation. Near homogeneity as judged by SDS-PAGE.
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.
|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.
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.
modificationsSome 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
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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.
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
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.
ab136649 has been referenced in 8 publications.
- Nihei Y et al. Poly-glycine-alanine exacerbates C9orf72 repeat expansion-mediated DNA damage via sequestration of phosphorylated ATM and loss of nuclear hnRNPA3. Acta Neuropathol 139:99-118 (2020). PubMed: 31642962
- Dubois ML et al. UBB pseudogene 4 encodes functional ubiquitin variants. Nat Commun 11:1306 (2020). PubMed: 32161257
- Logan NJ et al. Demethylation of ITGAV accelerates osteogenic differentiation in a blast-induced heterotopic ossification in vitro cell culture model. Bone 117:149-160 (2018). PubMed: 30219480
- 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 . 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). PubMed: 29974255
- Stopford MJ et al. C9ORF72 hexanucleotide repeat exerts toxicity in a stable, inducible motor neuronal cell model, which is rescued by partial depletion of Pten. Hum Mol Genet 26:1133-1145 (2017). ICC/IF ; Mouse . PubMed: 28158451
- Kohli JS et al. ETS1, nucleolar and non-nucleolar TERT expression in nevus to melanoma progression. Oncotarget 8:104408-104417 (2017). PubMed: 29262649
- Mori K et al. Reduced hnRNPA3 increases C9orf72 repeat RNA levels and dipeptide-repeat protein deposition. EMBO Rep 17:1314-25 (2016). PubMed: 27461252