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

Overview

• Product nameAnti-Fibrillarin antibody [38F3] - Nucleolar MarkerSee all Fibrillarin primary antibodies ...
• Description
  Mouse monoclonal [38F3] to Fibrillarin - Nucleolar Marker
• SpecificityThis 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.
• Tested applicationsFlow Cyt, IHC-P, ICC/IF, IHC-Fr, WB, ICC more details
• Species reactivity
  Reacts with: Mouse, Rat, Chicken, Human, Caenorhabditis elegans, Fruit fly (Drosophila melanogaster), Schizosaccharomyces pombe, African Green Monkey, Opossum
  Predicted to work with: Plants
• Immunogen

  Yeast nuclear preparation (S. cerevisiae). Hybridomas were screened by immunofluorescence on yeast cells and by western blotting on yeast protein homogenates (S. cerevisiae).

• Positive controlICC: Chicken embryonic fibroblast (CEF) cells, HEK 293 cells, SH-SY5Y cells
• General notesPfam number: PF01269. A reference below describes the characterization of D77, an antibody very similar but not identical to ab4566.

Properties

• FormLiquid
• Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
• Storage bufferPreservative: 10mM Sodium Azide
  Constituents: Tissue Culture Supernatant
• PurityTissue culture supernatant
• Purification notesSterile filtered.
• Clonality Monoclonal
• Clone number38F3
• IsotypeIgG1
• Research Areas
  • Tags & Cell Markers
  • Subcellular Markers
  • Nucleus
  • Nucleolus

  • Epigenetics and Nuclear Signaling
  • Chromatin Binding Proteins
  • DNA / RNA binding

Applications

Target

• FunctionInvolved in pre-rRNA processing. Utilizes the methyl donor S-adenosyl-L-methionine to catalyze 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.
• Sequence similaritiesBelongs 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 localizationNucleus > nucleolus. Fibrillar region of the nucleolus.

Target information above from: UniProt accession P22087 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 37662 Fruit fly (Drosophila melanogaster)
  • Entrez Gene: 2091 Human
  • Entrez Gene: 100044829 Mouse
  • Entrez Gene: 14113 Mouse
  • Entrez Gene: 292747 Rat
  • Omim: 134795 Human
  • SwissProt: Q9W1V3 Fruit fly (Drosophila melanogaster)
  • SwissProt: P22087 Human

  • SwissProt: P35550 Mouse
  • SwissProt: P22509 Rat
  • Unigene: 2821 Fruit fly (Drosophila melanogaster)
  • Unigene: 299002 Human
  • Unigene: 4595 Mouse
  • Unigene: 98803 Rat

  see all

• Alternative names
  34 kD nucleolar scleroderma antigen antibody34 kDa nucleolar scleroderma antigen antibody34-kD nucleolar scleroderma antigen antibody

  FBL antibodyFBRL_HUMAN antibodyFIB antibodyFIB1 antibodyFLRN antibodyNop1p antibodyRNA U3 small nucleolar interacting protein 1 antibodyRNU3IP1 antibodyrRNA 2' O methyltransferase fibrillarin antibodyrRNA 2'-O-methyltransferase fibrillarin antibody

  see all

Anti-Fibrillarin antibody [38F3] - Nucleolar Marker images

• 

  Immunocytochemistry/ Immunofluorescence - 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 - 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).

• 

  Immunocytochemistry/ Immunofluorescence - 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).
• 

  Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
  All lanes : Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566) at 1/2000 dilution (+ blocking buffer 1:1 in PBS + 0.1% Tween for 1 hour)
  
  Lane 1 : Lysate of the cytoplasmic protein fraction of HeLa cells
  Lane 2 : Lysate of the nuclear protein fraction of HeLa cells
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  An Alexa Fluor^®680-conjugated Goat anti-mouse polyclonal at 1/10000 dilution
  
  Performed under reducing conditions.
  
  Observed band size : 34 kDa (why is the actual band size different from the predicted?)
  

  This image is courtesy of an Abreview submitted by Svetlana Khoronenkova

  See Abreview

• 

  Flow Cytometry-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.
  
  
• 

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

  Image courtesy of an anonymous Abreview.

  See Abreview

Protocols

• Immunostaining of Cells in Tissue Culture