製品の概要

  • 製品名
    Anti-Fibrillarin antibody [38F3]
    Fibrillarin 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [38F3] to Fibrillarin
  • 特異性
    This clone was selected because it stains a single ~34kDa band on western blotting and shows a clear and strong punctate staining of yeast nuclei. It can therefore be used to identify nucleoli immunocytochemically. ab4566 was raised against yeast nuclear preps and the immunogen was identified as Nop1p, the yeast homolog of fibrillarin. Due to high aa homology the antibody should work with any specie possessing a nucleus, however this has not been tested.
  • アプリケーション
    適用あり: Flow Cyt, IHC-P, ICC/IF, IHC-Fr, WB, ICCmore details
  • 種交差性
    交差種: Mouse, Rat, Chicken, Human, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe, Zebrafish, African green monkey, Opossum
    交差が予測される動物種: Plants
  • 免疫原

    Tissue, cells or virus corresponding to Saccharomyces cerevisiae Fibrillarin.

    Yeast nuclear preparation (S. cerevisiae). Hybridomas were screened by immunofluorescence on yeast cells and by western blotting on yeast protein homogenates (S. cerevisiae).

  • ポジティブ・コントロール
    • ICC: Chicken embryonic fibroblast (CEF) cells, HEK 293 cells, SH-SY5Y cells
  • 特記事項

    Pfam number: PF01269. A reference below describes the characterization of D77, an antibody very similar but not identical to ab4566.

    Gives a much weaker signal in western blot compared to ab218846.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab4566 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
Flow Cyt 1/20.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-P Use at an assay dependent concentration. PubMed: 18528333
ICC/IF 1/1000 - 1/5000.

For IF of mammalian cells 1/500.

ab4566 is sensitive to aldehyde. Use a mild formalin fixation or acetone or methanol fixation as the target is nuclear.

IHC-Fr Use at an assay dependent concentration. PubMed: 18559082
WB 1/2000 - 1/10000. Detects a band of approximately 34 kDa. 1/2000 (cell lysates) - 1/10000 (nuclear fractions)(ECL). For other (non-ECL) western detection methods, 1/1000 - 1/5000. To detect mammalian fibrillarin on western blots by ECL, 1/500.
ICC Use at an assay dependent concentration.

ab4566 is sensitive to aldehyde. Use a mild formalin fixation or acetone or methanol fixation as the target is nuclear.

ターゲット情報

  • 機能
    S-adenosyl-L-methionine-dependent methyltransferase that has the ability to methylate both RNAs and proteins. Involved in pre-rRNA processing by catalyzing the site-specific 2'-hydroxyl methylation of ribose moieties in pre-ribosomal RNA. Site specificity is provided by a guide RNA that base pairs with the substrate. Methylation occurs at a characteristic distance from the sequence involved in base pairing with the guide RNA. Also acts as a protein methyltransferase by mediating methylation of 'Gln-105' of histone H2A (H2AQ104me), a modification that impairs binding of the FACT complex and is specifically present at 35S ribosomal DNA locus (PubMed:24352239).
  • 配列類似性
    Belongs to the methyltransferase superfamily. Fibrillarin family.
  • 翻訳後修飾
    By homology to other fibrillarins, some or all of the N-terminal domain arginines are modified to asymmetric dimethylarginine (DMA).
  • 細胞内局在
    Nucleus, nucleolus. Fibrillar region of the nucleolus.
  • Information by UniProt
  • 参照データベース
    see all
  • 別名
    • 34 kD nucleolar scleroderma antigen antibody
    • 34 kDa nucleolar scleroderma antigen antibody
    • Fbl antibody
    • FBRL_HUMAN antibody
    • FIB antibody
    • FIB1 antibody
    • FLRN antibody
    • Histone-glutamine methyltransferase antibody
    • Nop1p antibody
    • RNA U3 small nucleolar interacting protein 1 antibody
    • RNU3IP1 antibody
    • rRNA 2' O methyltransferase fibrillarin antibody
    • rRNA 2'-O-methyltransferase fibrillarin antibody
    see all

Anti-Fibrillarin antibody [38F3] 画像

  • Rat neurons and glial stained with mouse monoclonal to Fibrillarin (green) and with chicken antibody to neurofilament NF-H (red). Cells were counterstained with a fluorescent DNA probe (blue). Nuclear DNA is revealed with Hoechst dye (blue). Cultures were processed using our standard fixation and staining procedure (in protocol section).

  • Human neuroblastoma line SH-SY5Y stained with mouse monoclonal to Fibrillarin (green) and with chicken antibody to neurofilament NF-H (red) and counterstained with a fluorescent DNA probe (blue). Nuclear DNA is revealed with Hoechst dye (blue). The NF-H antibody was used at a dilution of 1/100000 and the fibrillarin monoclonal at 1/1000. Cultures were processed using standard fixation and staining procedure (in protocol section).

  • Image courtesy of an anonymous Abreview.

    Mouse embryonic fibroblast fractionation.
    Cytopl - cytoplasmic fraction.
    Nucl - nuclear fraction.
    20 µg of each loaded.

    ab4566 used at a 1/2000 dilution.
    The secondary used was an Alexa-Fluor 680 conjugated goat anti-mouse polyclonal used at a 1/10000 dilution.

    See Abreview

  • All lanes : Anti-Fibrillarin antibody [38F3] (ab4566) at 1/2000 dilution (incubated for 1 hour, diluted with Blocking buffer 1:1 in PBS+0.1%Tween)

    Lane 1 : cytoplasmic protein fraction of HeLa cells with LI-COR® Odyssey® Blocking Buffer, 45 minutes at room temperature at 50 %
    Lane 2 : nuclear protein fraction of HeLa cells with LI-COR® Odyssey® Blocking Buffer, 45 minutes at room temperature at 50 %

    Lysates/proteins at 20 µg per lane.

    Secondary
    AlexaFluor 680 goat anti-mouse at 1/10000 dilution

    Performed under reducing conditions.

    Additional bands at : 34 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 1 second

    Image is courtesy of Dr Svetlana Khoronenkova

    See Abreview

  • Overlay histogram showing HEK293 cells stained with ab4566 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab4566, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1](ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • ab staining Fibrillarin in Human melanoma A7 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton and blocked with 1% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (1/500 in 1% BSA) for 24 hours at 4°C. A FITC-conjugated Goat anti-mouse polyclonal (1/200) was used as the secondary antibody.

    See Abreview

  • High magnification view of human Hek293 cell nuclei stained with mouse monoclonal to fibrillarin (green), counterstained with a fluorescent DNA probe (blue). Nuclear DNA is revealed with Hoechst dye (blue). Cultures were processed using our standard fixation and staining procedure (in protocol section).

Anti-Fibrillarin antibody [38F3] (ab4566) 使用論文

This product has been referenced in:
  • Krejcí J  et al. Neural Differentiation in HDAC1-Depleted Cells Is Accompanied by Coilin Downregulation and the Accumulation of Cajal Bodies in Nucleoli. Stem Cells Int 2017:1021240 (2017). IHC ; Mouse . Read more (PubMed: 28337219) »
  • Shekari F  et al. Data for whole and mitochondrial proteome of human embryonic stem cells. Data Brief 13:371-376 (2017). WB ; Human . Read more (PubMed: 28664172) »

See all 91 Publications for this product

Product Wall

Filter by Application

Filter by Species

Filter by Ratings

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Embryonic Heart)
Permeabilization
Yes - Tween-20
Specification
Embryonic Heart
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Oct 21 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Heart)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
30 µg
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Oct 05 2016

Application
Immunoprecipitation
Immuno-precipitation step
Protein A/G
Sample
Human Cell lysate - whole cell (HeLa cells)
Specification
HeLa cells
Total protein in input
30 µg
Username

Abcam user community

Verified customer

投稿 Jan 15 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
Sample
Human Cell (Human primary fibroblasts)
Specification
Human primary fibroblasts
Permeabilization
Yes - 0.2% Triton-X100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Dec 17 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - nuclear (Mouse embryonic fibroblasts)
Loading amount
20 µg
Specification
Mouse embryonic fibroblasts
Gel Running Conditions
Reduced Denaturing (4-16% Tris-Gly gel)
Blocking step
LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
Username

Abcam user community

Verified customer

投稿 Jan 10 2012

Application
Immunocytochemistry/ Immunofluorescence
Sample
Caenorhabditis elegans Cell (intestinal nuclei)
Specification
intestinal nuclei
Fixative
Paraformaldehyde
Permeabilization
Yes - Sperm Salts Buffer, Triton X-100
Username

Abcam user community

Verified customer

投稿 Jun 03 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Fruit fly (Drosophila melanogaster) Cultured Cells (S2)
Specification
S2
Fixative
Paraformaldehyde
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Nov 26 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (HeLa cells)
Loading amount
20 µg
Specification
HeLa cells
Treatment
Fractionation of HeLa cells
Gel Running Conditions
Reduced Denaturing (10% Tris-Gly)
Blocking step
LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
Username

Dr. Svetlana Khoronenkova

Verified customer

投稿 Jul 23 2010

Application
Immunocytochemistry/ Immunofluorescence
Sample
Secale cereale Cell (Root-tip cells)
Specification
Root-tip cells
Fixative
Formaldehyde
Permeabilization
Yes - Triton X100, 0.1%
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 8%
Username

Dr. Manuela Silva

Verified customer

投稿 Apr 06 2010

Application
Immunocytochemistry/ Immunofluorescence
Sample
Arabidopsis suecica Cell (Leaf cell)
Specification
Leaf cell
Fixative
Formaldehyde
Permeabilization
Yes - Triton X100, 0.1%
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 8%
Username

Dr. Manuela Silva

Verified customer

投稿 Apr 06 2010

1-10 of 18 Abreviews

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

登録