Anti-Nucleolin antibody (ab22758)
- Datasheet
- References (62)
- Protocols
Overview
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Product nameAnti-Nucleolin antibody
See all Nucleolin primary antibodies -
DescriptionRabbit polyclonal to Nucleolin
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Host speciesRabbit
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Tested applicationsSuitable for: WB, IP, IHC-Fr, IHC-P, ICC/IFmore details
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Species reactivityReacts with: Mouse, Rat, Human
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Nucleolin.
Read Abcam's proprietary immunogen policy (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
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FormLiquid
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Storage instructionsShipped 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.
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Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Related Products
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
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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.
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Sequence similaritiesContains 4 RRM (RNA recognition motif) domains.
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Post-translational
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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Database links
- Entrez Gene: 4691 Human
- Entrez Gene: 17975 Mouse
- Entrez Gene: 25135 Rat
- Omim: 164035 Human
- SwissProt: P19338 Human
- SwissProt: P09405 Mouse
- SwissProt: P13383 Rat
- Unigene: 79110 Human
see all -
Alternative names
- C23 antibody
- FLJ45706 antibody
- MS1116 antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence - Anti-Nucleolin antibody (ab22758)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).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleolin antibody (ab22758)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. -
Western blot - Anti-Nucleolin antibody (ab22758)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
All lanes : 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. -
Immunoprecipitation - Anti-Nucleolin antibody (ab22758)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. -
Western blot - Anti-Nucleolin antibody (ab22758)Lane 1 : Marker
Lanes 2-5 : Anti-Nucleolin antibody (ab22758) at 1 µg/ml
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? -
Immunocytochemistry/ Immunofluorescence - Anti-Nucleolin antibody (ab22758)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.
Datasheets and documents
Protocols
References
This product has been referenced in:
- Zhang S et al. Dual radiosensitization and anti-STAT3 anti-proliferative strategy based on delivery of gold nanoparticle - oligonucleotide nanoconstructs to head and neck cancer cells. Nanotheranostics 2:1-11 (2018). Read more (PubMed: 29291159) »
- Telechea-Fernández M et al. New localization and function of calpain-2 in nucleoli of colorectal cancer cells in ribosomal biogenesis: effect of KRAS status. Oncotarget 9:9100-9113 (2018). Read more (PubMed: 29507677) »
