Overview

  • Product name

  • 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.

Properties

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.
  • 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

  • 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) 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.
  • 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.
  • 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.

  • 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.

References

This product has been referenced in:

  • Potapova TA  et al. Superresolution microscopy reveals linkages between ribosomal DNA on heterologous chromosomes. J Cell Biol 218:2492-2513 (2019). Read more (PubMed: 31270138) »
  • 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). Read more (PubMed: 28360124) »
See all 10 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Cat Cell (Crandell Rees feline kidney cells)
Permeabilization
Yes - Triton X-100
Specification
Crandell Rees feline kidney cells
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 37°C
Fixative
Paraformaldehyde

Delphine Acar

Verified customer

Submitted Jul 04 2018

Abreviews
Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Non-reduced Denaturing (7.5)
Sample
Human Cell lysate - whole cell (HeLa cells)
Specification
HeLa cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted May 28 2013

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

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
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
Application
Western blot
Sample
Human Cell lysate - whole cell (MRC5VA Lung Fibroblasts)
Loading amount
15 µg
Specification
MRC5VA Lung Fibroblasts
Gel Running Conditions
Reduced Denaturing (9%)
Blocking step
Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Feb 14 2011

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