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 Cyt, In-Cell ELISAmore details
Species reactivityReacts with: Human
Does not react with: Mouse, Rat, Cow
SU-DHL-1 Nuclei (Cell lysate nuclear fraction).
- HeLa and HDFn cells; HepG2, HDFn and SH-SY5Y whole cell lysates.
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: 0.36% HEPES, 0.88% Sodium chloride
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.
|In-Cell ELISA||Use a concentration of 1 µg/ml.|
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.
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) »