Overview

  • Product nameAnti-Fibrillarin [38F3] antibodySee all Fibrillarin primary antibodies ...
  • Description
    Mouse monoclonal [38F3] to Fibrillarin
  • SpecificityThis 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.
  • Tested applicationsFlow Cyt, IHC-P, ICC/IF, IHC-Fr, WB, ICC more details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Human, Caenorhabditis elegans, Fruit fly (Drosophila melanogaster), Schizosaccharomyces pombe, African Green Monkey, Opossum
    Predicted to work with: Plants
  • Immunogen

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

  • Positive control
    • ICC: Chicken embryonic fibroblast (CEF) cells, HEK 293 cells, SH-SY5Y cells
  • General notesPfam number: PF01269. A reference below describes the characterization of D77, an antibody very similar but not identical to ab4566.

Properties

Applications

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.

Application Notes
Flow Cyt 1/20.
IHC-P Use at an assay dependent dilution. PubMed: 18528333
ICC/IF 1/1000 - 1/5000. For IF of mammalian cells 1/500.
IHC-Fr Use at an assay dependent dilution. 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 dilution.

Target

  • FunctionInvolved in pre-rRNA processing. Utilizes the methyl donor S-adenosyl-L-methionine to catalyze 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.
  • Sequence similaritiesBelongs to the methyltransferase superfamily. Fibrillarin family.
  • Post-translational
    modifications
    By homology to other fibrillarins, some or all of the N-terminal domain arginines are modified to asymmetric dimethylarginine (DMA).
  • Cellular localizationNucleus > 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
  • Database links
    see all
  • Alternative names
    • 34 kD nucleolar scleroderma antigen antibody
    • 34 kDa nucleolar scleroderma antigen antibody
    • 34-kD nucleolar scleroderma antigen antibody
    • FBL antibody
    • FBRL_HUMAN antibody
    • FIB antibody
    • FIB1 antibody
    • FLRN 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 [38F3] antibody images

  • 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).

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

    Lane 1 : Lysate of the cytoplasmic protein fraction of HeLa cells
    Lane 2 : Lysate of the nuclear protein fraction of HeLa cells

    Lysates/proteins at 20 µg per lane.

    Secondary
    An Alexa Fluor®680-conjugated Goat anti-mouse polyclonal at 1/10000 dilution

    Performed under reducing conditions.

    Observed band size : 34 kDa (why is the actual band size different from the predicted?)

    This image is courtesy of an Abreview submitted by 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.

  • Image courtesy of an anonymous Abreview.

    See Abreview

References for Anti-Fibrillarin [38F3] antibody (ab4566)

This product has been referenced in:
  • Daneshvar K  et al. MicroRNA miR-308 regulates dMyc through a negative feedback loop in Drosophila. Biol Open 2:1-9 (2013). WB ; Fruit fly (Drosophila melanogaster) . Read more (PubMed: 23336071) »
  • Okoro DR  et al. Endogenous Human MDM2-C Is Highly Expressed in Human Cancers and Functions as a p53-Independent Growth Activator. PLoS One 8:e77643 (2013). Human . Read more (PubMed: 24147044) »

See all 35 Publications for this product

Product Wall

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
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Submitted Dec 17 2013

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...

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Thank you for calling Abcam earlier today.

I am sorry about the issues you have been having with ab4566 in western blot. As we discussed I am sending you a new vial of ab4566. Your new order number is *********.

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I would like to reassure you that these products are safe to use.

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.

Also, if y...

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I am sorry to hear of your customer's problems with this product. Could you please double check the purchase order provided as there does not seem to be an ab4566 associated with this PO.

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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
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Submitted Jan 10 2012

As requested, I have issued a free of charge replacement, ab4566, mouse fibrillarin antibody 38F3. To check the status of the order please contact our Customer Service team and reference this number. You should expect the antibody on Tuesday of next...

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Application Immunocytochemistry/ Immunofluorescence
Sample Caenorhabditis elegans Cell (intestinal nuclei)
Specification intestinal nuclei
Fixative Paraformaldehyde
Permeabilization Yes - Sperm Salts Buffer, Triton X-100
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Submitted Jun 03 2011

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