Overview

  • Product name
    Anti-Nucleolin antibody [3G4B2]
    See all Nucleolin primary antibodies
  • Description
    Mouse monoclonal [3G4B2] to Nucleolin
  • Host species
    Mouse
  • Specificity
    Recognizes human nucleolin, MW 100kD and an additional protein, MW 95kDa in HeLa nuclear extract. The clone number has been updated from (4i51) to (3G4B2) both clone numbers name the same antibody clone.
  • Tested applications
    Suitable for: EMSA, WB, IP, ELISA, ICCmore details
  • Species reactivity
    Reacts with: Dog, Human, Monkey
    Does not react with: Mouse, Rat
  • Immunogen

    Nuclear matrix extract from MDCK cells (dog).

  • Positive control
    • WB: HeLa nuclear extract. IP: MDCK total cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab14966 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
EMSA Use at an assay dependent dilution. PubMed: 16571724
WB 1/1000 - 1/5000. Detects a band of approximately 106 kDa (predicted molecular weight: 85 kDa). A weak non-specific band of ~60kDa is detected in human cell lines with lower antibody dilutions.
IP Use at an assay dependent dilution. (see Gotzmann et al reference).
ELISA Use at an assay dependent dilution.
ICC 1/10 - 1/100.

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

  • Anti-Nucleolin antibody [3G4B2] (ab14966) at 1/5000 dilution + Hela nuclear lysate

    Developed using the ECL technique.

    Predicted band size: 85 kDa



    HeLa nuclear extract was probed with anti-nucleolin (1:5000 dilution).

     

References

This product has been referenced in:
  • Wang Y  et al. PNRC accumulates in the nucleolus by interaction with B23/nucleophosmin via its nucleolar localization sequence. Biochim Biophys Acta : (2010). Read more (PubMed: 20888865) »
  • Hwang J  et al. Role of Scarf and its binding target proteins in epidermal calcium homeostasis. J Biol Chem 282:18645-53 (2007). IHC-P, IP . Read more (PubMed: 17470426) »
See all 4 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Question
Answer

I have been informed that the statement regarding IP for this antibody (which is the same as clone 3G4B2) comes from the reference Gotzmann J et al. Two-dimensional electrophoresis reveals a nuclear matrix-associated nucleolin complex of basic isoelectric point. Electrophoresis 18:2645-53 (1997). PubMed: 9527495 I found the article in PubMed, but the full length article was not available so I cannot quote a dilution value for IP, I'm very sorry I cannot help you more. I hope you will be able to access the paper and find this information, my apologies for the delay,

Read More

Answer

Thank you for your enquiry and patience. I was able to obtain the following information from the source of this antibody. Human nucleolin is consistently described as having two isoforms, with molecular weights of 95 kDa and 90 kDa. Ab14966 recognizes these isoforms and on a Western blot, there is a strong band at 95 kDa and a much fainter band at 90 kDa. This image will be added to our online datasheet. Here is the Western blot protocol as well: 1. Perform SDS-PAGE on a cell lysate sample (cell lysis buffer: 50mM Tris-HCl, pH7.4; 1% NP-40; 0.25% sodium deoxycholate; 150mM NaCl; 1mM EGTA; 1mM PMSF; 1ug/ml each aprotinin, leupeptin, pepstatin; 1mM Na3VO4; 1mM NaF) and transfer the proteins to nitrocellulose. Wash the blotted nitrocellulose twice with water. 2. Block the blotted nitrocellulose in freshly prepared PBS containing 3% nonfat dry milk (PBS-MLK) for 60 minutes at RT 3. Incubate the nitrocellulose with a 1:1000-1:5000 dilution of anti-Nucleolin diluted in freshly prepared PBS-MLK overnight with agitation at 4°C. 4. Wash the nitrocellulose twice with water. 5. Incubate the nitrocellulose in the secondary reagent of choice (a goat anti-mouse HRP conjugated IgG, 1:5000 dilution was used) in PBS-MLK for 1.0 hours at RT with agitation. 6. Wash the nitrocellulose with water twice. 7. Wash the nitrocellulose in PBS-0.05% Tween 20 for 15 minutes. 8. Rinse the nitrocellulose in 4-5 changes of water with a final wash in water for 2 hours at RT with agitation. 9. Use detection method of choice (enhanced chemiluminescence was used).

Read More

Answer

Hello Abcam, This antibody is an unpurified cell culture supernant, so the antibody concentration can not be determined against the background of other proteins. The lack of a known concentration is compensated for by stated specific dilutions for suggested applications. The antibody is recommended for immunoprecipitation citing a specificic reference: Gotzmann, J., et al., Electrophoresis 18: 2645–2653, 1997. An epitope determination has not been performed.

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