Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
![Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)](https://www.abcam.com/ps/products/4/ab4566/Images/ab4566_1.jpg "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
Overview
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Product name
Anti-Fibrillarin antibody [38F3] - Nucleolar Marker
See all Fibrillarin primary antibodies -
Description
Mouse monoclonal [38F3] to Fibrillarin - Nucleolar Marker -
Host species
Mouse -
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. 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.
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Tested applications
Suitable for: Flow Cyt, ICC/IF, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Plants
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Immunogen
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). -
Positive control
- ICC: HEK-293 cells and SH-SY5Y cells; WB: HEK-293, C6, NIH-3T3 nuclear fractions
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General notes
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.
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Properties
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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: 0.065% Sodium azide
Constituent: Tissue culture supernatant -
Concentration information loading... -
Purity
Tissue culture supernatant -
Purification notes
Sterile filtered. -
Clonality
Monoclonal -
Clone number
38F3 -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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 | Abreviews | Notes |
|---|---|---|
| Flow Cyt |
1/20.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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| 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. |
| Notes |
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Flow Cyt
1/20. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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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. |
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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. |
Target
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Function
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). -
Sequence similarities
Belongs to the methyltransferase superfamily. Fibrillarin family. -
Post-translational
modificationsBy 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. - Information by UniProt
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Database links
- 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
see all -
Alternative names
- 34 kD nucleolar scleroderma antigen antibody
- 34 kDa nucleolar scleroderma antigen antibody
- Fbl antibody
see all
Images
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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).
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![Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)](https://www.abcam.com/ps/products/4/ab4566/Images/ab4566-342869-AntiFibrillarinantibody38F3helahuman.jpg)
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).
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![Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)](https://www.abcam.com/ps/products/4/ab4566/Images/ab4566-342868-AntiFibrillarinantibody38F3westernblot.jpg)
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)](https://www.abcam.com/ps/products/4/ab4566/Images/ab4566_2.jpg)
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).
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![Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)](https://www.abcam.com/ps/products/4/ab4566/Images/ab4566-9284-anti-fibrillarin-antibody-38f3-western-blot.jpg)
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. -
![Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)](https://www.abcam.com/ps/products/4/ab4566/Images/ab4566-262968-anti-fibrillarin-antibody-38f3-western-blot.jpg)
Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)Image is courtesy of Dr Svetlana KhoronenkovaAll 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
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![Flow Cytometry - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)](https://www.abcam.com/ps/products/4/ab4566/Images/ab4566-9285-anti-fibrillarin-antibody-38f3-flow-cytometry.jpg)
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)](https://www.abcam.com/ps/products/4/ab4566/Images/ab4566-262967-anti-fibrillarin-antibody-38f3-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)Image is courtesy of Cesar Camachoab 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.
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![Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)](https://www.abcam.com/ps/products/4/ab4566/Images/ab4566_3.jpg)
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).
Datasheets and documents
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SDS download
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Datasheet download
References (198)
ab4566 has been referenced in 198 publications.
- Sun Z et al. LIN28 coordinately promotes nucleolar/ribosomal functions and represses the 2C-like transcriptional program in pluripotent stem cells. Protein Cell 13:490-512 (2022). PubMed: 34331666
- Sepich-Poore C et al. The METTL5-TRMT112 N6-methyladenosine methyltransferase complex regulates mRNA translation via 18S rRNA methylation. J Biol Chem 298:101590 (2022). PubMed: 35033535
- Tagliaferro M et al. Nucleolar localization of the ErbB3 receptor as a new target in glioblastoma. BMC Mol Cell Biol 23:13 (2022). PubMed: 35255831
- Matsumori H et al. Ribosomal protein L5 facilitates rDNA-bundled condensate and nucleolar assembly. Life Sci Alliance 5:N/A (2022). PubMed: 35321919
- Li N et al. A sphingolipid-mTORC1 nutrient-sensing pathway regulates animal development by an intestinal peroxisome relocation-based gut-brain crosstalk. Cell Rep 40:111140 (2022). PubMed: 35905721