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Tags & Cell Markers Subcellular Markers Nucleus Nucleolus

Anti-Fibrillarin抗体[38F3] - Nucleolar Marker (ab4566)

  • Datasheet
  • SDS
Reviews (24)Q&A (18)References (176)

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Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
  • Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
  • Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
  • Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
  • Flow Cytometry - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)

Key features and details

  • Mouse monoclonal [38F3] to Fibrillarin - Nucleolar Marker
  • Suitable for: Flow Cyt, ICC/IF, WB
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG1

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概述

  • 产品名称

    Anti-Fibrillarin抗体[38F3] - Nucleolar Marker
    参阅全部 Fibrillarin 一抗
  • 描述

    小鼠单克隆抗体[38F3] to Fibrillarin - Nucleolar Marker
  • 宿主

    Mouse
  • 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. 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.

  • 经测试应用

    适用于: Flow Cyt, ICC/IF, WBmore details
  • 种属反应性

    与反应: Mouse, Rat, Human
    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: HEK-293 cells and SH-SY5Y cells; WB: HEK-293, C6, NIH-3T3 nuclear fractions
  • 常规说明

    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.

    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

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • 存储溶液

    Preservative: 0.065% Sodium azide
    Constituent: Tissue culture supernatant
  • Concentration information loading...
  • 纯度

    Tissue culture supernatant
  • 纯化说明

    Sterile filtered.
  • 克隆

    单克隆
  • 克隆编号

    38F3
  • 同种型

    IgG1
  • 研究领域

    • Tags & Cell Markers
    • Subcellular Markers
    • Nucleus
    • Nucleolus
    • Epigenetics and Nuclear Signaling
    • Chromatin Binding Proteins
    • DNA / RNA binding

相关产品

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • Recombinant Protein

    • Recombinant Human Fibrillarin protein (ab99443)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab4566于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
Flow Cyt
1/20.

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

ICC/IF (9)
1/100.

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.

WB (10)
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.

说明
Flow Cyt
1/20.

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

ICC/IF
1/100.

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.

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.

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.
  • Target information above from: UniProt accession P22087 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 2091 Human
    • Entrez Gene: 102643269 Mouse
    • Entrez Gene: 14113 Mouse
    • Entrez Gene: 292747 Rat
    • Omim: 134795 Human
    • SwissProt: P22087 Human
    • SwissProt: P35550 Mouse
    • SwissProt: P22509 Rat
    • Unigene: 299002 Human
    • Unigene: 4595 Mouse
    • Unigene: 98803 Rat
    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

图片

  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
    Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)

    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
    Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)

    ICC analysis of HeLa cells stained with mouse monoclonal to Fibrillarin (green) and with chicken antibody to vimentin (red) and counterstained with a fluorescent DNA probe (blue). Nuclear DNA is revealed with DAPI(blue). The vimentin antibody was used at a dilution of 1/1000 and the fibrillarin monoclonal at 1/100. Cultures were processed using standard fixation and staining procedure (in protocol section).

  • Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
    Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
    All lanes : Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566) at 1/500 dilution

    Lane 2 : C6 cytosol fraction
    Lane 3 : C6 nuclear fraction
    Lane 4 : HEK-293 cytosol fraction
    Lane 5 : HEK-293 nuclear fraction
    Lane 6 : NIH-3T3 cytosol fraction
    Lane 7 : NIH-3T3 nuclear fraction

    Observed band size: 37 kDa why is the actual band size different from the predicted?

  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
    Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)

    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).

  • Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
    Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)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

  • Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
    Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)Image is courtesy of Dr Svetlana Khoronenkova
    All lanes : Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (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
    All lanes : 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

    See Abreview

  • Flow Cytometry - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
    Flow Cytometry - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
    Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)Image is courtesy of Cesar Camacho

    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

  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
    Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
    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).

实验方案

  • Immunostaining of Cells in Tissue Culture

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (176)

发表研究结果有使用 ab4566?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab4566 被引用在 176 文献中.

  • Kim ES  et al. C9orf72-associated arginine-rich dipeptide repeats induce RNA-dependent nuclear accumulation of Staufen in neurons. Hum Mol Genet 30:1084-1100 (2021). PubMed: 33783499
  • Villacampa A  et al. From Spaceflight to Mars g-Levels: Adaptive Response of A. Thaliana Seedlings in a Reduced Gravity Environment Is Enhanced by Red-Light Photostimulation. Int J Mol Sci 22:N/A (2021). PubMed: 33477454
  • Hasygar K  et al. Coordinated control of adiposity and growth by anti-anabolic kinase ERK7. EMBO Rep 22:e49602 (2021). PubMed: 33369866
  • Weeks SE  et al. Inhibiting ß-catenin disables nucleolar functions in triple-negative breast cancer. Cell Death Dis 12:242 (2021). PubMed: 33664239
  • Zhu Q  et al. Rcl1 depletion impairs 18S pre-rRNA processing at the A1-site and up-regulates a cohort of ribosome biogenesis genes in zebrafish. Nucleic Acids Res 49:5743-5759 (2021). PubMed: 34019640
View all Publications for this product

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Immunocytochemistry/ Immunofluorescence abreview for Anti-Fibrillarin antibody [38F3] - Nucleolar Marker

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Thyroid cancer cell lines (Cal62 and TPC1))
Permeabilization
Yes - Triton 0.5%
Specification
Thyroid cancer cell lines (Cal62 and TPC1)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°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.

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提交于 Jun 08 2022

Western blot abreview for Anti-Fibrillarin antibody [38F3] - Nucleolar Marker

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (MEF)
Gel Running Conditions
Reduced Denaturing (9%)
Loading amount
15 µg
Specification
MEF
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°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.

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提交于 Jan 04 2019

Immunocytochemistry abreview for Anti-Fibrillarin antibody [38F3] - Nucleolar Marker

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Mouse Cell (fibroblast)
Permeabilization
Yes - methanol
Specification
fibroblast
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Formaldehyde
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.

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提交于 Dec 05 2018

Immunoprecipitation abreview for Anti-Fibrillarin antibody [38F3] - Nucleolar Marker

Average
Abreviews
Abreviews
abreview image
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (U2OS cells)
Total protein in input
500 µg
Immuno-precipitation step
Protein A/G
Specification
U2OS cells
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.

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提交于 Mar 15 2018

Western blot abreview for Anti-Fibrillarin antibody [38F3] - Nucleolar Marker

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS cells)
Gel Running Conditions
Reduced Denaturing (4-12% Gradient Gel)
Loading amount
50 µg
Specification
U2OS cells
Blocking step
Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°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.

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提交于 Mar 06 2018

Western blot abreview for Anti-Fibrillarin antibody [38F3] - Nucleolar Marker

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (MEF cells)
Gel Running Conditions
Reduced Denaturing (4-12% Gradient Gel)
Loading amount
50 µg
Specification
MEF cells
Blocking step
Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°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.

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提交于 Feb 14 2018

Immunohistochemistry (Frozen sections) abreview for Anti-Fibrillarin antibody [38F3] - Nucleolar Marker

Excellent
Abreviews
Abreviews
abreview image
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
Read More
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提交于 Oct 21 2016

Western blot abreview for Anti-Fibrillarin antibody [38F3] - Nucleolar Marker

Good
Abreviews
Abreviews
abreview image
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
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.

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提交于 Oct 05 2016

Immunoprecipitation abreview for Anti-Fibrillarin antibody [38F3] - Nucleolar Marker

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

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提交于 Jan 15 2015

Immunocytochemistry/ Immunofluorescence abreview for Anti-Fibrillarin antibody [38F3] - Nucleolar Marker

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

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提交于 Dec 17 2013

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研究类别(A-Z)
  • 肿瘤研究
  • 心血管研究
  • 细胞生物学
  • 发育生物学
  • 染色质及细胞核信号转导研究
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