Key features and details
- Mouse monoclonal [38F3] to Fibrillarin - Nucleolar Marker
- Suitable for: Flow Cyt, ICC/IF, WB
- Reacts with: Mouse, Rat, Chicken, Human, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe, Zebrafish, African green monkey, Opossum
- Isotype: IgG1
Product nameAnti-Fibrillarin antibody [38F3] - Nucleolar Marker
See all Fibrillarin primary antibodies
DescriptionMouse monoclonal [38F3] to Fibrillarin - Nucleolar Marker
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.
Tested applicationsSuitable for: Flow Cyt, ICC/IF, WBmore details
Species reactivityReacts with: Mouse, Rat, Chicken, Human, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe, Zebrafish, African green monkey, Opossum
Predicted to work with: Plants
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).
- ICC: Chicken embryonic fibroblast (CEF) cells, HEK-293 cells, SH-SY5Y cells; WB: HEK-293, C6, NIH-3T3 nuclear fractions
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.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferPreservative: 0.065% Sodium azide
Constituent: Tissue culture supernatant
Concentration information loading...
PurityTissue culture supernatant
Purification notesSterile filtered.
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.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
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.|
FunctionS-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).
Sequence similaritiesBelongs to the methyltransferase superfamily. Fibrillarin family.
modificationsBy 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.
- Information by UniProt
- 34 kD nucleolar scleroderma antigen antibody
- 34 kDa nucleolar scleroderma antigen antibody
- Fbl antibody
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).
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).
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?
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).
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.
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.
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
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.
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).
ab4566 has been referenced in 143 publications.
- 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