Anti-Fibrillarin抗体[38F3] (ab4566)

概述

  • 产品名称Anti-Fibrillarin抗体[38F3]
    参阅全部 Fibrillarin 一抗
  • 描述
    小鼠单克隆抗体[38F3] to Fibrillarin
  • 特异性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.
  • 经测试应用适用于: Flow Cyt, IHC-P, ICC/IF, IHC-Fr, WB, ICCmore details
  • 种属反应性
    与反应: Mouse, Rat, Chicken, Human, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe, Zebrafish, African Green Monkey, Opossum
    预测可用于: 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).

  • 阳性对照
    • ICC: Chicken embryonic fibroblast (CEF) cells, HEK 293 cells, SH-SY5Y cells
  • 常规说明Pfam number: PF01269. A reference below describes the characterization of D77, an antibody very similar but not identical to ab4566.

性能

应用

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.

应用 Ab评论 说明
Flow Cyt 1/20.
IHC-P Use at an assay dependent concentration. PubMed: 18528333
ICC/IF 1/1000 - 1/5000. For IF of mammalian cells 1/500.
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.

靶标

  • 功能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).
  • 序列相似性Belongs to the methyltransferase superfamily. Fibrillarin family.
  • 翻译后修饰By homology to other fibrillarins, some or all of the N-terminal domain arginines are modified to asymmetric dimethylarginine (DMA).
  • 细胞定位Nucleus, nucleolus. Fibrillar region of the nucleolus.
  • Information by UniProt
  • 数据库链接
    see all
  • 别名
    • 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] 图像

  • 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)参考文献

This product has been referenced in:
  • Guan Y  et al. Phosphorylation of Def Regulates Nucleolar p53 Turnover and Cell Cycle Progression through Def Recruitment of Calpain3. PLoS Biol 14:e1002555 (2016). IHC ; Zebrafish . Read more (PubMed: 27657329) »
  • Walz K  et al. Characterization of ANKRD11 mutations in humans and mice related to KBG syndrome. Hum Genet 134:181-90 (2015). Read more (PubMed: 25413698) »

See all 52 Publications for this product

Product Wall

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
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提交于 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
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提交于 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
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提交于 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

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Verified customer

提交于 Dec 17 2013

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Expiration date: 21 March 2012


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The anti-Fibrillarin antibody [38F3] is a tissue culture supernatant and is not purified. Other proteins will be within the solution and this can give it an off white colour. ...

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Since you can see expected positivity along with what appears to be non specifici cytosolic staining, I would suggest diluting the primary 2 - 4 fold, which will hopefully reduce non specific background while retaining specificity.

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