Anti-Nucleolin antibody ab22758 is a rabbit polyclonal antibody that is used in Nucleolin western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.
- Tried and trusted by researchers since 2006
IgG
Rabbit
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Liquid
Polyclonal
IHC-P | IP | WB | ICC/IF | IHC-Fr | |
---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Expected |
Mouse | Expected | Expected | Tested | Expected | Expected |
Rat | Expected | Expected | Tested | Expected | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1 µg/mL | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1 µg/mL | Notes - |
Species Rat | Dilution info 1 µg/mL | Notes - |
Species Human | Dilution info 1 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Human | Dilution info Use at an assay dependent concentration. | Notes - |
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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.
Nucleolin, Protein C23, NCL
Anti-Nucleolin antibody ab22758 is a rabbit polyclonal antibody that is used in Nucleolin western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.
- Tried and trusted by researchers since 2006
IgG
Rabbit
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Liquid
Polyclonal
Affinity purification Immunogen
Replenishment batches of our polyclonal antibody, ab22758 are tested in WB. Previous batches were additionally validated in ICC/IF, IHC-Fr, IHC-P and IP. These applications are still expected to work and are covered by our Abpromise guarantee. You may also be interested in our alternative recombinant antibody, ab129200.
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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This supplementary information is collated from multiple sources and compiled automatically.
Nucleolin also known as C23 is a multifunctional protein mainly found in the nucleolus with a molecular weight of approximately 100 kDa. It plays a mechanical role in ribosomal RNA (rRNA) synthesis and ribosome biogenesis. Nucleolin is widely expressed in proliferating cells including HeLa culture and shows higher expression in cancer cells compared to normal cells. The nucleolin protein is involved in various nucleolar processes like the modulation of chromatin structure and nucleocytoplasmic transport.
The nucleolin molecular weight protein contributes to regulating cellular processes such as chromatin remodeling rRNA transcription and mRNA stability. Nucleolin often forms complexes with various other proteins and nucleic acids enhancing its functionality within the cell nucleus. Its participation in processes like DNA replication and cell proliferation shows the central role nucleolin plays at the cellular level. It influences cellular responses by binding directly to specific RNA or DNA sequences.
The nucleolin protein engages in major cellular pathways such as the RNA processing and p53 signaling pathways. Through the RNA processing pathway it interacts with proteins like fibrillarin to assist in rRNA maturation. Moreover within the p53 signaling pathway nucleolin regulates the activity of the tumor suppressor protein p53 impacting cell cycle progression and apoptosis. These interactions highlight nucleolin's role in maintaining cellular homeostasis and response to stress.
The nucleolin protein often shows altered expression in cancers and cardiovascular diseases. In cancer nucleolin's high expression levels correlate with tumor progression and metastasis making it a potential target for cancer therapies. In cardiovascular diseases nucleolin interacts with proteins like VEGF to influence angiogenesis which affects disease progression. Understanding nucleolin's involvement in these diseases provides insight into potential therapeutic targets and interventions.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
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) (Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) ab99697).
Band: 100kDa: Nucleolin.
All lanes: Immunoprecipitation - Anti-Nucleolin antibody (ab22758)
Predicted band size: 76 kDa
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: Western blot - Anti-Nucleolin antibody (ab22758) at 1 µg/mL
Lane 1: MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg
Lane 2: Brain (Mouse) Tissue Lysate at 10 µg
Lane 3: Pancreas (Mouse) Tissue Lysate at 10 µg
Lane 4: PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg
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, 60 kDa
All lanes: Western blot - Anti-Nucleolin antibody (ab22758)
Lanes 2 and 4: Western blot - Jurkat whole cell lysate (Jurkat whole cell lysate ab7899) at 20 µg
Lanes 3 and 5: Western blot - A-431 whole cell lysate (A-431 whole cell lysate ab7909) at 20 µg
All lanes: 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
ab22758 staining nucleolin in HeLa cells treated with Triptolide from Tripterygium wilfordii (Triptolide from Tripterygium wilfordii, Anti-inflammatory agent 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 Triptolide from Tripterygium wilfordii, Anti-inflammatory agent 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 (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/250 dilution was used as the secondary antibody.
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