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Immunocytochemistry/ Immunofluorescence - Fibrillarin antibody - Nucleolar Marker (ab5821)
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Immunofluorescence - Fibrillarin antibody - Nucleolar Marker (ab5821)
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Western blot - Fibrillarin antibody - Nucleolar Marker (ab5821)
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Product Name
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Fibrillarin antibody - Nucleolar Marker
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See all Fibrillarin - Nucleolar Marker antibodies (5)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to Fibrillarin - Nucleolar Marker
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Immunogen
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Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Fibrillarin.
(Peptide available as ab13745.)
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Reacts with
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Hu, Ms, Dm, Xl
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Does not react with
(species key)
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Rat
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Specificity
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This antibody detects a band at close to 34kDa in all species tested. The band can be completely blocked with the immuising peptide in all cases - this is very strong evidence that the antibody is recognising fibrillarin.
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Tested applications
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ICC/IF, IHC-P, IP, WB
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Abreviews
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Application notes
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Recommended dilutions
ICC: 1/100.
ICC/IF: Use at an assay dependent dilution.
IF: 1/50.
IHC-P: 1/100.
IP: Use at an assay dependent dilution (PMID 19430468).
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 39 kDa (predicted molecular weight: 34 kDa). Can be blocked with Fibrillarin peptide (ab13745).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Cellular localization
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Nuclear; fibrillar region of the nucleolus.
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Research areas
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Chromatin and Nuclear Signaling >> Chromatin Binding Proteins >> DNA / RNA binding
Tags & Cell Markers >> Subcellular Markers >> Nucleus >> Nucleolus
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Relevance
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Nop1p / Fibrillarin was originally identified as a nucleolar protein of bakers 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 homologues to the other sequences throughout all of the fibrillarin domain. The structure of this molecule has been determined and shown to consist of 2 extended b-sheets flanked by 4 a-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.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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Batch dependent within range: 0.40 - 0.80 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this Fibrillarin antibody - Nucleolar Marker is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for Fibrillarin antibody - Nucleolar Marker images, references, products related to ab5821 and other tools.
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Fibrillarin antibody - Nucleolar Marker images:
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 Immunocytochemistry/ Immunofluorescence - Fibrillarin antibody - Nucleolar Marker (ab5821)
ICC/IF image of ab5821 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab5821, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
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Immunofluorescence - Fibrillarin antibody - Nucleolar Marker (ab5821)
Immunofluorescent imaging of human cells (U2OS) with ab5821 reveals highly specific localisation to the dense fibrillar component (DFC) of the nucleolus associated with the initial ribosomal RNA (rRNA) precursor. The nucleolar protein fibrillarin is located primarily in the DFC. Blue is hoechst staining of the nucleus, green is ab5821 used at 1/100, merge image demonstrates exclusively nuclear localisation.
IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour. Primary antibody used at 1/100 in 5% milk / 0.2% TWEEN for one hour, secondary antibody Alexa 488 for 30 minutes. All blocking and incubation steps carried out at 37 degrees C. Luke Hughes-Davies and Rhiannon Jade, Gurdon Institute, Cambridge, UK
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Western blot - Fibrillarin antibody - Nucleolar Marker (ab5821)
All lanes : Fibrillarin antibody - Nucleolar Marker (ab5821) at 1 µg/ml
Lane 1 : Hela nuclear lysate
Lane 2 : Hela whole cell lysate
Lane 3 : A431 whole cell lysate
Lane 4 : Jurkat whole cell lysate
Lane 5 : 293T whole cell lysate
Lane 6 : Hela nuclear lysate with Fibrillarin peptide (ab13745) at 1 µg/ml
Lane 7 : Hela whole cell lysate with Fibrillarin peptide (ab13745) at 1 µg/ml
Lane 8 : A431 whole cell lysate with Fibrillarin peptide (ab13745) at 1 µg/ml
Lane 9 : Jurkat whole cell lysate with Fibrillarin peptide (ab13745) at 1 µg/ml
Lane 10 : 293T whole cell lysate with Fibrillarin peptide (ab13745) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L (HRP) (ab6721) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 34 kDa
Exposure time : 10 seconds
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Fibrillarin antibody - Nucleolar Marker (ab5821)
ab5821 staining human liver tissue sections by IHC-P. Sections were formaldehyde fixed, permeabilized in Triton-X and subjected to heat mediated antigen retrieval prior to blocking with 6% BSA for 2 hours at 23°C. The primary antibody was diluted 1/100 and incubated with the sample for 16 hours at 5°C. ab6717 was used as the secondary antibody. This image is courtesy of an Abreview submitted by Dr Tom Donndelinger
See Abreview
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References for Fibrillarin antibody - Nucleolar Marker (ab5821)
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This product has been used in: (two most recent references)
Sutton GM et al. Central nervous system melanocortin-3 receptors are required for synchronizing metabolism during entrainment to restricted feeding during the light cycle. FASEB J 24:862-72 (2010). WB; Mouse. PubMed: 19837866
Miron MJ et al. Localization and importance of the adenovirus E4orf4 protein during lytic infection. J Virol 83:1689-99 (2009). ICC/IF; Human. PubMed: 19073741
See all 20 publication references for this product.
If you publish research using ab5821 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to Fibrillarin
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