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

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Overview

Product name

Anti-Fibrillarin antibody [38F3] - Nucleolar Marker
See all Fibrillarin products (5) ...

Description

Mouse monoclonal [38F3] to Fibrillarin - Nucleolar Marker

Specificity

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.

Tested applications

Flow Cyt, in situ hybridisation, IHC-P, ICC/IF, IHC-Fr, WB, IF, ICCmore details

Cross 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 notes

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

Properties

Form

Liquid

Storage instructions

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

Storage buffer

Preservative: 10mM Sodium Azide
Constituents: Tissue Culture Supernatant

Purity

Tissue culture supernatant

Purification notes

Sterile filtered.

Clonality

Monoclonal

Clone number

38F3

Isotype

IgG1

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

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

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

Applications

Show applications key

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.

Target

Function

Involved 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 similarities

Belongs 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 localization

Nucleus > nucleolus. Fibrillar region of the nucleolus.

Target information above from: UniProt accessionP22087 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).

Information by UniProt

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
  • RNU3IP1 antibody
  • rRNA 2' O methyltransferase fibrillarin antibody
  • rRNA 2'-O-methyltransferase fibrillarin antibody
see all

Database links

see all

Anti-Fibrillarin antibody [38F3] - Nucleolar Marker images:

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

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

Immunocytochemistry/ Immunofluorescence - 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).

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

Immunocytochemistry/ Immunofluorescence - 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).

  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 1/2000 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?)


Blocking Step: 50% Li-COR® Odyssey® Bloc for 45 minutes at room temperature

Cell fractionation was performed as described in Woodhouse BC et al DNA Repair (Amst). 2008: 7(6):932-40.

This image is courtesy of an Abreview submitted by Svetlana Khoronenkova

See Abreview

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

Flow Cytometry-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.

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

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

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.

Image courtesy of an anonymous Abreview.

See Abreview

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

This product has been referenced in:

  • Baxley RMet al. The role of the Suppressor of Hairy-wing insulator protein in Drosophila oogenesis. Dev Biol 356:398-410 (2011). IHC-Fr; Fruit fly (Drosophila melanogaster).Read more (PubMed: 21651900) »
  • Basu Aet al. Requirement of rRNA methylation for 80S ribosome assembly on a cohort of cellular internal ribosome entry sites. Mol Cell Biol 31:4482-99 (2011).Read more (PubMed: 21930789) »

See all 28 publications for this product

Publishing research using ab4566? Please let us know so that we can cite the reference in this datasheet

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