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  1. Link

    nucleolin-antibody-ab70493.pdf

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Tags & Cell Markers Subcellular Markers Nucleus Nucleolus
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Anti-Nucleolin antibody (ab70493)

  • Datasheet
Reviews (3)Q&A (3)References (11)

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Western blot - Anti-Nucleolin antibody (ab70493)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleolin antibody (ab70493)
  • Immunoprecipitation - Anti-Nucleolin antibody (ab70493)
  • Immunocytochemistry/ Immunofluorescence - Anti-Nucleolin antibody (ab70493)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleolin antibody (ab70493)

Key features and details

  • Rabbit polyclonal to Nucleolin
  • Suitable for: IHC-P, ICC/IF, WB, IP
  • Reacts with: Mouse, Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-Nucleolin antibody
    See all Nucleolin primary antibodies
  • Description

    Rabbit polyclonal to Nucleolin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, ICC/IF, WB, IPmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rabbit, Horse, Guinea pig, Cow, Dog, Pig, Chimpanzee, Rhesus monkey, Gorilla, African green monkey, Orangutan, Elephant
  • Immunogen

    Synthetic peptide corresponding to a region between residue 550 and the C-terminus (residue 710) of human Nucleolin

  • Positive control

    • Whole cell lysates from HeLa cells, 293T cells and mouse NIH3T3 cells.
  • General notes

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 6.8
    Preservative: 0.09% Sodium azide
    Constituents: 0.1% BSA, Tris buffered saline
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Subcellular Markers
    • Nucleus
    • Nucleolus
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Synthesis
    • Other
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • Translation
    • Ribosome
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • RNA Processing
    • Other

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • HeLa whole cell lysate (ab29545)
    • NIH 3T3 whole cell lysate (ab7179)
  • Recombinant Protein

    • Recombinant Human Nucleolin protein (His tag) (ab241527)

Applications

Our Abpromise guarantee covers the use of ab70493 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
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.
WB 1/2000 - 1/10000. Detects a band of approximately 100 kDa (predicted molecular weight: 77 kDa).
IP Use at 2-5 µg/mg of lysate.

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.
  • Target information above from: UniProt accession P19338 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4691 Human
    • Entrez Gene: 17975 Mouse
    • Entrez Gene: 100174230 Orangutan
    • Omim: 164035 Human
    • SwissProt: P19338 Human
    • SwissProt: P09405 Mouse
    • SwissProt: Q5RF26 Orangutan
    • Unigene: 79110 Human
    • Unigene: 154378 Mouse
    • Unigene: 474153 Mouse
    see all
  • Alternative names

    • C23 antibody
    • FLJ45706 antibody
    • MS1116 antibody
    • NCL antibody
    • Nucl antibody
    • NUCL_HUMAN antibody
    • Nucleolin antibody
    • Protein C23 antibody
    see all

Images

  • Western blot - Anti-Nucleolin antibody (ab70493)
    Western blot - Anti-Nucleolin antibody (ab70493)
    All lanes : Anti-Nucleolin antibody (ab70493) at 0.02 µg/ml

    Lane 1 : Whole cell lysate from HeLa cells at 50 µg
    Lane 2 : Whole cell lysate from HeLa cells at 15 µg
    Lane 3 : Whole cell lysate from HeLa cells at 5 µg
    Lane 4 : Whole cell lysate from 293T cells at 50 µg
    Lane 5 : Whole cell lysate from NIH3T3 cells at 50 µg

    Predicted band size: 77 kDa
    Observed band size: 100 kDa
    why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleolin antibody (ab70493)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleolin antibody (ab70493)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma (left) and mouse squamous cell carcinoma (right) tissues labelling Nucleolin with ab70493 at 1/1000 (0.2µg/ml). Detection: DAB.
  • Immunoprecipitation - Anti-Nucleolin antibody (ab70493)
    Immunoprecipitation - Anti-Nucleolin antibody (ab70493)
    Detection of Human Nucleolin by Immunoprecipitation in Whole cell lysate from HeLa cells (1 mg for IP, 20% of IP loaded), using ab70493 at 3 µg/mg lysate (Lane 1). Lane 2 represents rabbit IgG IP control. Subsequent Western blot detection of nucleolin was performed using ab70493 at 1 µg/ml.
  • Immunocytochemistry/ Immunofluorescence - Anti-Nucleolin antibody (ab70493)
    Immunocytochemistry/ Immunofluorescence - Anti-Nucleolin antibody (ab70493)
    ICC/IF image of ab70493 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab70493, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleolin antibody (ab70493)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleolin antibody (ab70493)
    IHC image of ab70493 staining in human normal lymph node 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 ab70493, 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.

    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.

Protocols

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
  • References (11)

    Publishing research using ab70493? Please let us know so that we can cite the reference in this datasheet.

    ab70493 has been referenced in 11 publications.

    • Potapova TA  et al. Superresolution microscopy reveals linkages between ribosomal DNA on heterologous chromosomes. J Cell Biol 218:2492-2513 (2019). PubMed: 31270138
    • Turi Z  et al. Perturbation of RNA Polymerase I transcription machinery by ablation of HEATR1 triggers the RPL5/RPL11-MDM2-p53 ribosome biogenesis stress checkpoint pathway in human cells. Cell Cycle 17:92-101 (2018). PubMed: 29143558
    • Evsyukov V  et al. Genetic mutations linked to Parkinson's disease differentially control nucleolar activity in pre-symptomatic mouse models. Dis Model Mech 10:633-643 (2017). PubMed: 28360124
    • Duncan FE  et al. Age-associated dysregulation of protein metabolism in the mammalian oocyte. Aging Cell 16:1381-1393 (2017). ICC/IF ; Mouse . PubMed: 28994181
    • Lista MJ  et al. Nucleolin directly mediates Epstein-Barr virus immune evasion through binding to G-quadruplexes of EBNA1 mRNA. Nat Commun 8:16043 (2017). WB . PubMed: 28685753
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a question

    1-3 of 3 Q&A

    Question

    Dear Abcam technical support,
    Thank you for offering your help in testing the different lots of this antibody with the mouse lung. Certainly, I would like to see if your lab can find out any lot of this antibody that can produce the apical staining that I showed you in my previous email. We have found another nucleolin antibody that give us some of the staining we wanted but still didn't work as well as we expected. Also, since this staining pattern was published on Nature and your antibody was referenced, we would still want to produce consistent results with the same ab22758 antibody. Please inform me when you have the results and I'm looking forward to see if we can get the apical staining back.
    Really appreciate for your help and please let me know if I might be helpful to provide you additional information to assist your test on the antibody.

    Read More

    Abcam community

    Verified customer

    Asked on Jun 05 2012

    Answer

    Thank you for your reply.
    I have let the lab know that they should re-test the antibody with mouse lung samples, and I will let you know as soon as I have the data. I will also send on the previous IHC images once I have access to them.
    Please let me know if there is anything else that we can do. I am sorry again for the batch variability, but hopefully this will be resolved shortly.
    Have a great day.

    Read More

    Abcam Scientific Support

    Answered on Jun 05 2012

    Question


    Sorry that it took me this long to reply your email, I was occupied with my work and I need some time to gather some information that you requested about my inquiry. I had taken two pictures on the IHC staining that I did on mouse lungs with ab22758 nucleolin antibody in separate time. The first one was done in January 2010, lot number was 724414, and this slide shows the staining that we expected on the apical aspect of the bronchioles. I have added some red arrows to point out the apical staining, which looks like someone outlines the lining using a marker. The second slide was done early this year, lot number GR8557-2 and the staining was done using the same protocol as the previous, and all the apical staining were not detected. This apical staining should appear in the mouse lung bronchioles. I have added some red arrows on this second slide too to indicate where the apical staining supposed to be. They were not picked up by the antibody but instead stained in blue by the hematoxylin.
    I hope this will be sufficient information for you to investigate what might be the problem between these two lots and please contact me anytime if you need additional information. Thank you again for your time in this matter.

    Read More

    Abcam community

    Verified customer

    Asked on Jun 01 2012

    Answer

    Thank you for your reply.
    I forwarded the images and information to my colleagues in the lab, and we have only tested this antibody in tonsil tissue. My colleague who specifically works with this antibody is out of the office until next office, but we'll be able to take a look at the previous data once she returns.
    The lab is also offering to test a couple of the current lots in mouse lung to see if one works better; would you like us to do this? I'm not sure what the time frame would be to get the results, but I would send them to you once they're available.
    Please let me know if you would be interested in this and I will set it up with the lab. I look forward to hearing from you. Have a great weekend!

    Read More

    Abcam Scientific Support

    Answered on Jun 01 2012

    Question

    Dear abcam technical support:

    I just called minutes ago about the ab22758 antibody and I’ve received great supports from your operator. The reason I called in for technical support is because we would like to run the same test on new batch of experimental samples but we couldn’t get the apical staining back anymore as they’ve published on the paper. This problem started after we got the new lot of this antibody and it has been missing the apical staining on the mouse lungs. Even performing the same staining protocol on the samples that worked before, which were used for the paper, with this new lot antibody, the apical staining is not showing.

    We had called in to asked for replacement vial but it didn’t solve the problem. As you might recalled from our conversation record, we’ve also tried ab70493 and that didn’t help as well. We’ve also been asking if there’s any new lot of the ab22758 that might be coming soon but seems like you still have a lot of vials in stock of this lot.

    From our conversation today over the phone, seems like the only thing we can find out is if there’s anything unusual about the QC on this lot. Would there be anything else that might led to the variation of this lot compare to the previous? I hope you can help us find out if there’s any way that we can get the apical staining back since this is the only antibody that we had consistent success with the old lots. We’ve tried all other antibodies that target Nucleolin but nothing give us the staining we wanted.

    Thank you in advance for your help

    Read More

    Abcam community

    Verified customer

    Asked on Apr 23 2012

    Answer

    Thank you for your call today and for your email.
    I apologize again for the batch variability of ab22758, and I have a couple of additional questions. Do you know which lot number was used previously to produce the apical staining? If not, do you have an order or PO number (I can look up the lot number)? This will be helpful for comparing QC data.
    Also, do you have any images of the most recent staining with this lot?
    I look forward to hearing from you and I'm hopeful that we can resolve this issue. Please let me know if you have any questions or if there is anything else that we can do for you.

    Read More

    Abcam Scientific Support

    Answered on Apr 23 2012

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