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Mouse Monoclonal Fibrillarin antibody. Nucleolus marker. Suitable for Flow Cyt, WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 223 publications. Immunogen corresponding to Cell preparation containing NOP1 protein.


Images

Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (AB4566), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (AB4566), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (AB4566), expandable thumbnail
  • Flow Cytometry - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (AB4566), expandable thumbnail
  • Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (AB4566), expandable thumbnail

Publications

Key facts

Isotype

IgG1

Host species

Mouse

Storage buffer

Preservative: 0.065% Sodium azide
Constituents: Tissue culture supernatant

Form

Liquid

Clonality

Monoclonal

Immunogen

  • Cell preparation containing NOP1 protein. The exact immunogen used to generate this antibody is proprietary information. Database link P15646

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
Flow CytWBICC/IF
Human
Tested
Tested
Tested
Mouse
Expected
Tested
Expected
Rat
Expected
Tested
Tested
Plants
Predicted
Predicted
Predicted

Tested
Tested

Species

Human

Dilution info

1/20

Notes

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Plants

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/2000.00000 - 1/10000.00000

Notes

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.

Species

Rat

Dilution info

1/2000.00000 - 1/10000.00000

Notes

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.

Species

Human

Dilution info

1/2000.00000 - 1/10000.00000

Notes

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.

Predicted
Predicted

Species

Plants

Dilution info

-

Notes

-

Tested
Tested

Species

Rat

Dilution info

1/100

Notes

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.

Species

Human

Dilution info

1/100

Notes

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.

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Plants

Dilution info

-

Notes

-

Associated Products

Select an associated product type

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Target data

Function

S-adenosyl-L-methionine-dependent methyltransferase that has the ability to methylate both RNAs and proteins (PubMed:24352239, PubMed:30540930, PubMed:32017898). Involved in pre-rRNA processing by catalyzing the site-specific 2'-hydroxyl methylation of ribose moieties in pre-ribosomal RNA (PubMed:30540930). Site specificity is provided by a guide RNA that base pairs with the substrate (By similarity). Methylation occurs at a characteristic distance from the sequence involved in base pairing with the guide RNA (By similarity). Probably catalyzes 2'-O-methylation of U6 snRNAs in box C/D RNP complexes (PubMed:32017898). U6 snRNA 2'-O-methylation is required for mRNA splicing fidelity (PubMed:32017898). 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, PubMed:30540930). Part of the small subunit (SSU) processome, first precursor of the small eukaryotic ribosomal subunit. During the assembly of the SSU processome in the nucleolus, many ribosome biogenesis factors, an RNA chaperone and ribosomal proteins associate with the nascent pre-rRNA and work in concert to generate RNA folding, modifications, rearrangements and cleavage as well as targeted degradation of pre-ribosomal RNA by the RNA exosome (PubMed:34516797).

Targets

NOP1

Alternative names

Recommended products

Mouse Monoclonal Fibrillarin antibody. Nucleolus marker. Suitable for Flow Cyt, WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 223 publications. Immunogen corresponding to Cell preparation containing NOP1 protein.

Key facts

Isotype

IgG1

Form

Liquid

Clonality

Monoclonal

Immunogen
  • Cell preparation containing NOP1 protein. The exact immunogen used to generate this antibody is proprietary information. Database link P15646
Clone number

38F3

Purity

Tissue culture supernatant

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.

Concentration
Loading...
Purification notes

Sterile filtered.

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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 Anti-Fibrillarin antibody [4A4] ab218846.

Abcam is leading the way to address reproducibility in scientific research with our highly validated recombinant monoclonal and recombinant multiclonal antibodies. Search & select one of Abcam's thousands of recombinant alternatives to eliminate batch-variability and unnecessary animal use.

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9 product images

  • Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566), expandable thumbnail
    Image courtesy of an anonymous customer review.

    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.

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

    Predicted band size: 33 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566), expandable thumbnail

    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566), expandable thumbnail

    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).

  • Flow Cytometry - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566), expandable thumbnail

    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) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1](Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control 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), expandable thumbnail

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

    All lanes: Western blot - 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

    Predicted band size: 33 kDa

    Observed band size: 37 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566), expandable thumbnail

    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), expandable thumbnail
    Image is courtesy of Dr Svetlana Khoronenkova

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

    All lanes: Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566) at 1/2000 dilution

    Lane 1: cytoplasmic protein fraction of HeLa cells at 20 µg

    Lane 2: nuclear protein fraction of HeLa cells at 20 µg

    Secondary

    All lanes: AlexaFluor 680 goat anti-mouse at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 33 kDa

    Observed band size: 34 kDa

    Exposure time: 1s

  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566), expandable thumbnail
    Image is courtesy of Cesar Camacho

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

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566), expandable thumbnail

    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).

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