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Anti-Nucleolin 抗体 (ab70493)

Reviews (3)Q&A (3)References (17)
Western blot - Anti-Nucleolin antibody (ab70493)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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, ICC, WB, IP
  • Reacts with: Mouse, Human
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-Nucleolin antibody
    Nucleolin 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to Nucleolin

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: IHC-P, ICC/IF, ICC, WB, IPmore details

  • 種交差性

    交差種: Mouse, Human
    交差が予測される動物種: Rabbit, Horse, Guinea pig, Cow, Dog, Pig, Chimpanzee, Rhesus monkey, Gorilla, African green monkey, Orangutan, Elephant

  • 免疫原

    Synthetic peptide within Human Nucleolin aa 550 to the C-terminus (C terminal). The exact immunogen sequence used to generate this antibody is proprietary information. If additional detail on the immunogen is needed to determine the suitability of the antibody for your needs, please contact our Scientific Support team to discuss your requirements.
    Database link: P19338

  • ポジティブ･コントロール

    • WB: HeLa, HEK-293T and NIH/3T3 whole cell lysate. IHC-P: Human ovarian carcinoma and mouse squamous cell carcinoma tissue. Human normal lymph node tissue. Mouse cerebellum tissue. IP: HeLa whole cell lysate. ICC/IF: HeLa cells.

  • 特記事項

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab70493の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
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 (1)
Use at an assay dependent concentration.
ICC
Use a concentration of 5 µg/ml.
WB (2)
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.
特記事項
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 at an assay dependent concentration.
ICC
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.

ターゲット情報

  • 機能

    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.

  • 配列類似性

    Contains 4 RRM (RNA recognition motif) domains.

  • 翻訳後修飾

    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.

  • 細胞内局在

    Nucleus > nucleolus. Cytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

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

画像

  • 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.
  • 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 mouse cerebellum tissue labelling Nucleolin with ab70493 at 1/100 (2 µg/ml), followed by goat anti-rabbit IgG-heavy and light chain cross-adsorbed antibody DyLight® 594 used at 1/100 dilution.

  • 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.

参考文献 (17)

ab70493 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab70493 は 17 報の論文で使用されています。

  • Potapova TA  et al. Distinct states of nucleolar stress induced by anticancer drugs. Elife 12:N/A (2023). PubMed: 38099650
  • Moudry P  et al. RNA-interference screen for p53 regulators unveils a role of WDR75 in ribosome biogenesis. Cell Death Differ 29:687-696 (2022). PubMed: 34611297
  • Mirra S  et al. Ubiquitin Specific Protease USP48 Destabilizes NF-κB/p65 in Retinal Pigment Epithelium Cells. Int J Mol Sci 23:N/A (2022). PubMed: 36077078
  • Angrand G  et al. Type I arginine methyltransferases are intervention points to unveil the oncogenic Epstein-Barr virus to the immune system. Nucleic Acids Res 50:11799-11819 (2022). PubMed: 36350639
  • Goldson TM  et al. Nucleolin mediates the binding of cancer cells to L-selectin under conditions of lymphodynamic shear stress. Am J Physiol Cell Physiol 318:C83-C93 (2020). PubMed: 31644306
View all Publications for this product

レビューと Q&A

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1-6 of 6 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-Nucleolin antibody

 Excellent
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Delphine Acar

Verified customer

投稿 Jul 04 2018

Western blot

 Average
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 May 28 2013

Western blot abreview for Anti-Nucleolin antibody

 Excellent
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Feb 14 2011

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
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

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
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

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