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
  • アプリケーション適用あり: IHC-P, ICC/IF, 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 corresponding to a region between residue 550 and the C-terminus (residue 710) of human Nucleolin

  • ポジティブ・コントロール
    • Whole cell lysates from HeLa cells, 293T cells and mouse NIH3T3 cells.

製品の特性

アプリケーション

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.

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

Anti-Nucleolin antibody 画像

  • 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.
  • 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?)
  • 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.

Anti-Nucleolin antibody (ab70493) 使用論文

This product has been referenced in:
  • Guzman-Ayala M  et al. Chd1 is essential for the high transcriptional output and rapid growth of the mouse epiblast. Development 142:118-27 (2015). Read more (PubMed: 25480920) »
  • Fulka H & Langerova A The maternal nucleolus plays a key role in centromere satellite maintenance during the oocyte to embryo transition. Development 141:1694-704 (2014). ICC/IF . Read more (PubMed: 24715459) »

See all 4 Publications for this product

Product Wall

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

投稿 May 28 2013

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.
Pleas...

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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, bu...

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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 ...

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Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

投稿 Feb 14 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"