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

Reviews (23)Q&A (18)References (143)

Key features and details

  • Mouse monoclonal [38F3] to Fibrillarin - Nucleolar Marker
  • Suitable for: Flow Cyt, ICC/IF, WB
  • Reacts with: Mouse, Rat, Chicken, Human, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe, African green monkey, Opossum
  • Isotype: IgG1

製品の概要

  • 製品名

    Anti-Fibrillarin antibody [38F3] - Nucleolar Marker
    Fibrillarin 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [38F3] to Fibrillarin - Nucleolar Marker

  • 由来種

    Mouse

  • 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, ICC/IF, WBmore details

  • 種交差性

    交差種: Mouse, Rat, Chicken, Human, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe, 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; WB: HEK-293, C6, NIH-3T3 nuclear fractions

  • 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.
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.

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

画像

  • 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).

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

    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).

参考文献 (143)

ab4566 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。

ab4566 は 143 報の論文で使用されています。

  • Wang H  et al. Mitochondrial and calcium perturbations in rat CNS neurons induce calpain-cleavage of Parkin: Phosphatase inhibition stabilizes pSer65Parkin reducing its calpain-cleavage. Biochim Biophys Acta Mol Basis Dis 1865:1436-1450 (2019). PubMed: 30796971
  • Martinez-Macias MI  et al. FUS (fused in sarcoma) is a component of the cellular response to topoisomerase I-induced DNA breakage and transcriptional stress. Life Sci Alliance 2:N/A (2019). PubMed: 30808650
  • Ren X  et al. Maintenance of Nucleolar Homeostasis by CBX4 Alleviates Senescence and Osteoarthritis. Cell Rep 26:3643-3656.e7 (2019). PubMed: 30917318
  • Chai W  et al. HMGB1-mediated autophagy regulates sodium/iodide symporter protein degradation in thyroid cancer cells. J Exp Clin Cancer Res 38:325 (2019). PubMed: 31331356
  • Jezek J  et al. Mitochondrial translocation of cyclin C stimulates intrinsic apoptosis through Bax recruitment. EMBO Rep 20:e47425 (2019). PubMed: 31385392
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1-10 of 41 Abreviews or Q&A

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

 Excellent
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
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投稿 Jan 04 2019

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

 Good
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
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投稿 Dec 05 2018

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

 Average
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
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Verified customer

投稿 Mar 15 2018

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

 Excellent
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
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Verified customer

投稿 Mar 06 2018

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

 Excellent
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
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Abcam user community

Verified customer

投稿 Feb 14 2018

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

 Excellent
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
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Verified customer

投稿 Oct 21 2016

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

 Good
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
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Abcam user community

Verified customer

投稿 Oct 05 2016

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

 Good
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
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Abcam user community

Verified customer

投稿 Jan 15 2015

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

 Excellent
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
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Abcam user community

Verified customer

投稿 Dec 17 2013

Question

We would like to test Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566) for zebrafish using Western Blot. We would like to apply for Abreivew discount. Please advise.

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Answer

DISCOUNT CODE: ******
Expiration date: 21 March 2012


I am very pleased to hear you would like to accept our offer and test ab4566 in Zebrafish. This code will give you the value of ab4566 off your next order before the expiration date. To redeem this offer, please submit an Abreview for Zebrafish and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: https://www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: https://www.abcam.com/abtrial.

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