Fibrillarin Antibody (38F3)

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] - 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.

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

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] - Human SH-SY5Y cells stained with NB300-269, showing prominent specular nucleolar staining. The nuclei are counter stained with DAPI (blue), so these spots appear very pale blue. Neurofilament heavy protein was stained with NB300-217 (red).

Immunocytochemistry/Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] - Fibrillarin immunofluorescence in fibroblasts. Image from 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, Ye
• Applications: WB, Flow, ICC/IF, IHC, IHC-Fr, 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:
  • Western Blot 1:500
  • Flow Cytometry 1:20-1:400
  • Immunocytochemistry/Immunofluorescence 1:100
  • Immunohistochemistry 1:100
  • Immunohistochemistry-Frozen
  • Immunohistochemistry Whole-Mount
  • Knockdown Validated
• 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).
• 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 reported in scientific literature (PMID: 24722283 and 27588463 respectively).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Preservative: 10mM 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)(12)

Publications using NB300-269

• 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)
• Tiku V. Small Nucleoli and Reduced Ribosomal Biogenesis are Hallmarks of Longevity Thesis 2017 (ICC/IF, Drosophila)
• 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)
• 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)
• 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)
• 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)

Reviews for Fibrillarin Antibody (NB300-269) (3)

Average Rating: 5

(Based on 3 reviews)

We have 3 reviews tested in 3 species: Human, C. elegans, Other.

reviewed by: Varnesh Tiku WB C. elegans 01/09/2019

Summary

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

reviewed by: Anony- mous ICC Human 06/09/2014

Summary

• Application: Immunocytochemistry
• Species: Human
• Special Applications: 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).
• File: View PDF

reviewed by: Anonymous WB Other 04/24/2014

Summary

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

Blocking

• Blocking Details: blocked in 5% Normal Goat serum/0.3% Triton X-100 in PBS, 1 hour, room temperature

Primary Anitbody

• Dilution Ratio: 1:500, 4C, overnight, in 1X PBS/1% BSA/0.3% Triton X-100

Secondary Antibody

• Secondary Description: goat anti mouse alexafluor 488 conjugated
• Secondary Manufacturer Cat#: cell signaling 4408
• Secondary Concentration: 1:1000

Details

• Fixation Details: Fixed in PFA, Blocked in Normal Goat serum containing Triton X-100, Incubated overnight 1:100 in 1% BSA
• Wash Description: 1X PBS, 3 washes 5 minutes each

Bioinformatics

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