Fibrillarin Antibody (38F3)

Immunocytochemistry/Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] - High magnification confocal image of HeLa cells stained with fibrillarin antibody, dilution 1:100 from concentrated tissue culture media (Green), and costained with chicken vimentin pAb (Red), dilution 1:1,000. Nuclear DNA is revealed with the DAPI stain (Blue). The fibrillarin antibody shows strong staining of nucleoli in the nucleus, while the vimentin antibody reveals cytoplasmic intermediate filaments.

Western Blot: Fibrillarin Antibody (38F3) [NB300-269] - Analysis of lysates of cell fractions probed with Fibrillarin antibody, dilution 1:500 (Green): [1] protein standard, [2] C6 cytosol, [3] C6 nuclear, [4] HEK293 cytosol, [5] HEK293 nuclear, [6] NIH-3T3 cytosol and [7] NIH-3T3 nuclear fractions. The band at 37kDa corresponds to the fibrillarin protein detected exclusively in the nuclear fractions.

Immunocytochemistry/Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] - The nuclei of hth mutants have dispersed rDNA foci and ectopic nucleoli and produce more rRNA. A) Wild type nuclei stained with anti-Fibrillarin (red) and a specific probe that recognizes the rDNA genomic region (green). B) hth mutant nuclei stained as in A. The mutant nuclei exhibit an increased number of nucleoli with different sizes. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0120662), licensed under a CC-BY licence.

Immunohistochemistry-Paraffin: Fibrillarin Antibody (38F3) [NB300-269] - IHC staining of Fibrillarin in human ovarian cancer using DAB with hematoxylin counterstain.

Immunocytochemistry/Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] - Mouse embryonic fibroblast fibrillarin stained with fibrillarin antibody and Alexa-594 secondary antibody. ICC/IF image submitted by a verified customer review.

Flow Cytometry: Fibrillarin Antibody (38F3) [NB300-269] - Fibrillarin antibody was tested at 1:400 in HEK293 cells using an Alexa Fluor 488 secondary (shown in green) alongside unstained cells (shown in red).

Western Blot: Fibrillarin Antibody (38F3) [NB300-269] - Western blot analysis of Fibrillarin expression on yeast protein extracts using NB300-269.

Immunocytochemistry/Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] - Fibrillarin immunofluorescence in fibroblasts. ICC/IF image submitted by a verified customer review.

Immunocytochemistry/Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] - IF Confocal analysis of MCF7 cells using Fibrillarin antibody (NB300-269, 1:5). An Alexa Fluor 488-conjugated Goat to mouse IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).

• Reactivity: Hu, Mu, Rt, Po, Bv, Ce, Ch, Dr, Eq, Pl, Y, Ye
• Applications: WB, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IHC-WhMt, KD
• Clone: 38F3
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

Fibrillarin Antibody (38F3) Summary

• Immunogen: Yeast nuclear preparation (S. cerevisiae).
• Localization: Nucleus > nucleolus. Note: Fibrillar region of the nucleolus.
• Marker: Nucleolar Marker
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: FBL
• Purity: Affinity purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:20-1:400
  • Immunocytochemistry/Immunofluorescence 1:500-1:1000
  • Immunohistochemistry Whole-Mount
  • Immunohistochemistry 1:500-1:1000
  • Immunohistochemistry-Frozen 1:500-1:1000
  • Immunohistochemistry-Paraffin 1:500-1:1000
  • Knockdown Validated
  • Western Blot 1:1000-1:5000
• Application Notes: This Fibrillarin Antibody (38F3) is useful for Western blot, Immunocytochemistry/Immunofluorescence, Flow Cytometry and Immunohistochemistry on both paraffin-embedded and frozen sections. In WB a band can be seen at approximately 34.5 kDa. In ICC/IF, this antibody shows prominent specular nucleolar staining.
  
  Use in IHC-WhMt and IHC-Fr reported in scientific literature (PMID: 24722283 and 21539824 respectively). Use in Immunohistochemistry-paraffin reported in scientific literature (PMID: 23542174).
• Theoretical MW: 34.5 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  Fibrillarin Overexpression Lysate

Reactivity Notes

C. elegans and plant reactivity reported in scientific literature (PMID: 24722283 and 27588463 respectively). Use in S. pombe reported in scientific literature (PMID:31855181).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Supplied as concentrated hybridoma cell culture media.
• Preservative: 5mM Sodium Azide
• Concentration: 1 mg/ml
• Purity: Affinity purified

Alternate Names for Fibrillarin Antibody (38F3)

• EC 2.1.1,34-kD nucleolar scleroderma antigen
• EC 2.1.1.-
• EC 2.1.1.37
• FIB
• FIB1,34 kDa nucleolar scleroderma antigen
• fibrillarin
• FLRNrRNA 2'-O-methyltransferase fibrillarin
• RNA, U3 small nucleolar interacting protein 1
• RNU3IP1

Background

Fibrillarin / Nop1p, was originally identified as a nucleolar protein of baker's yeast, Saccharomyces cerevisiae (accession P15646). The Nop1p protein is essential for yeast viability and is localized in the nucleoli. The human homologue of Nop1p is Fibrillarin (accession P22087), a component of the nucleolar small nuclear ribonucleoprotein (snRNP) particle. The human Fibrillarin gene is located on chromosome 19 (19q13.1). Fibrillarin proteins have been cloned and sequenced from several other species (mouse, accession P35550, Xenopus accession P22232, C. elegans accession Q22053, and S. pombe accession P35551). The N terminal 80 amino acids contain multiple copies based on the peptide RGG, and the remaining 240 amino acids consist of the Fibrillarin domain. A Fibrillarin homologue has also been identified in the genome of the archean Methanococcus (accession NC_000909). This protein lacks the RGG rich N-terminal extension but is clearly homologous to the other sequences throughout the entire Fibrillarin domain. The structure of this molecule has been determined and shown to consist of 2 extended beta-sheets flanked by 4 alpha-helixes. Patients with the autoimmune disease scleroderma often have strong circulating autoantibodies to a 34kDa protein which was subsequently found to be Fibrillarin. Fibrillarin is an excellent marker for the nucleolus.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Fibrillarin Antibody (NB300-269)(20)

Publications using NB300-269

• Hibino E, Ichiyama Y, Tsukamura A et al. Bex1 is essential for ciliogenesis and harbours biomolecular condensate-forming capacity BMC biology Feb 10 2022 [PMID: 35144600] (ICC/IF)
• Chen P, Chen Y, Chu T et al. Nucleolar control by a non apoptotic p53 caspases deubiquitinylase axis promotes resistance to bacterial infection FASEB J Nov 29 2019 (WB, C. elegans)
• Wu T, Nance J, Chu F, Fazzio T Characterization of R-loop-interacting proteins in embryonic stem cells reveals roles in ribosomal RNA processing and gene expression Molecular & Cellular Proteomics Aug 1 2021 [PMID: 34478875]
• Procida T, Friedrich T, Jack APM, et al. JAZF1, A Novel p400/TIP60/NuA4 Complex Member, Regulates H2A.Z Acetylation at Regulatory Regions International journal of molecular sciences Jan 12 2021 [PMID: 33445503] (ICC/IF, Human)
• Sural S, Liang CY, Wang FY et al. HSB-1/HSF-1 pathway modulates histone H4 in mitochondria to control mtDNA transcription and longevity Sci Adv Oct 1 2020 [PMID: 33087356]
• Begik O, Lucas MC, Liu H et al. Integrative analyses of the RNA modification machinery reveal tissue- and cancer-specific signatures Genome Biol. May 7 2020 [PMID: 32375858] (IHC-Fr, Mouse)
• Hsiung K, Liu K, Tsai T et al. Defects in CISD-1, a mitochondrial iron-sulfur protein, lower glucose level and ATP production in Caenorhabditis elegans Biomedical Journal Feb 1 2020 [PMID: 32200954] (WB, C. elegans)
• Wong IN, Neo JP, Oehler J et al. The Fml1-MHF complex suppresses inter-fork strand annealing in fission yeast Elife Dec 19 2019 [PMID: 31855181] (WB, S. pombe)
• Tiku V, Kew C, Mehrotra P et al. Nucleolar fibrillarin is an evolutionarily conserved regulator of bacterial pathogen resistance. Nat Commun Sep 6 2018 [PMID: 30190478] (WB, C. elegans)
• Buchwalter A, Hetzer MW. Nucleolar expansion and elevated protein translation in premature aging Nat Commun 2017 Aug 30 [PMID: 28855503] (ICC/IF, Human)
• Tiku V, Jain C, Raz Y et al. Small nucleoli are a cellular hallmark of longevity Nat Commun 2016 Aug 30 [PMID: 28853436] (WB, C. elegans)
• Robaszkiewicz E, Idziak-Helmcke D, Tkacz MA et al. The arrangement of Brachypodium distachyon chromosomes in interphase nuclei. J. Exp. Bot. Oct 1 2016 [PMID: 27588463] (IHC, PLANT)
• Mitrea DM, Cika JA, Guy CS et al. Nucleophosmin integrates within the nucleolus via multi-modal interactions with proteins displaying R-rich linear motifs and rRNA Elife Mar 1 2016 [PMID: 26836305] (ICC/IF, Mouse)
• Itatani Y, Sonoshita M, Kakizaki F et al. Characterization of Aes nuclear foci in colorectal cancer cells. J. Biochem. 2015 Jul 29 [PMID: 26229111] (ICC/IF, Human)
  
  Details:
  Fibrillarin antibody was used for ICC/IF analysis of HCT116 cells overexpressing AcGFP-tagged Aes (AcGFP-Aes) and TLE1 (Fig. 1B).
• Zaballos MA, Cantero W, Azpiazu N. The TALE Transcription Factor Homothorax Functions to Assemble Heterochromatin during Drosophila Embryogenesis. PLoS ONE. 2015 Mar 21 [PMID: 25794008] (IHC, Drosophila)
• Lee CC, Tsai YT, Kao CW et al. Mutation of a Nopp140 gene dao-5 alters rDNA transcription and increases germ cell apoptosis in C. elegans. Cell Death Dis 2014 Apr 11 [PMID: 24722283] (IHC-WhMt, C. elegans)
• Zillner K, Filarsky M, Rachow K et al. Large-scale organization of ribosomal DNA chromatin is regulated by Tip5 Nucleic Acids Res 2013 Apr 10 [PMID: 23580549] (ICC/IF, Human)
• Beyret E, Lin H. Pinpointing the expression of piRNAs and function of the PIWI protein subfamily during spermatogenesis in the mouse Dev Biol 2011 Jul 15 [PMID: 21539824] (IHC-Fr, Mouse)
• Tiku V. Small Nucleoli and Reduced Ribosomal Biogenesis are Hallmarks of Longevity Thesis 2017 (ICC/IF, Drosophila)
• Su H, Xu T, Ganapathy S et al. Elevated snoRNA biogenesis is essential in breast cancer Oncogene 2013 Apr 1 [PMID: 23542174] (WB, IHC-P, Human)

Reviews for Fibrillarin Antibody (NB300-269) (4)

Average Rating: 5

(Based on 4 reviews)

We have 4 reviews tested in 4 species: Human, Mouse, C. elegans, Other.

reviewed by: Ahmed Mostafa Immunofluorescence Mouse 06/14/2021

Summary

• Application: Immunofluorescence
• Sample Tested: fibroblasts,Mouse embryonic fibroblasts
• Species: Mouse
• Lot: 9A-1147725

reviewed by: Verified Customer Western Blot C. elegans 01/09/2019

Summary

• Application: Western Blot
• Sample Tested: 4% PFA-PBS fixed frozen sections from murine brain,Tissue Lysates
• Species: C. elegans
• Lot: 38F3

reviewed by: Verified Customer Immunocytochemistry Human 06/09/2014

Summary

• Application: Immunocytochemistry
• Species: Human

reviewed by: Jennifer Fox Western Blot Other 04/24/2014

Summary

• Application: Western Blot
• Sample Tested: fibroblasts
• Species: Other
• Lot: 92412

Bioinformatics

• Gene Symbol: FBL
• Entrez:
  • 2091(Human)
  • 14113(Mouse)
  • 292747(Rat)