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  1. Link

    fibrillarin-antibody-nucleolar-marker-ab5821.pdf

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Tags & Cell Markers Subcellular Markers Nucleus Nucleolus
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Anti-Fibrillarin antibody - Nucleolar Marker (ab5821)

  • Datasheet
  • SDS
Reviews (22)Q&A (6)References (158)

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Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody - Nucleolar Marker (ab5821)
  • Western blot - Anti-Fibrillarin antibody - Nucleolar Marker (ab5821)
  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody - Nucleolar Marker (ab5821)

Key features and details

  • Rabbit polyclonal to Fibrillarin - Nucleolar Marker
  • Suitable for: ICC/IF, WB
  • Reacts with: Mouse, Human, Drosophila melanogaster
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-Fibrillarin antibody [EPR10823(B)] - Nucleolar Marker (ab166630)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-Fibrillarin antibody - Nucleolar Marker
    See all Fibrillarin primary antibodies
  • Description

    Rabbit polyclonal to Fibrillarin - Nucleolar Marker
  • Host species

    Rabbit
  • Specificity

    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.
  • Tested applications

    Suitable for: ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human, Drosophila melanogaster
    Predicted to work with: Xenopus laevisDoes not react with: Rat
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • ICC/IF: HeLa cells
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Subcellular Markers
    • Nucleus
    • Nucleolus
    • Epigenetics and Nuclear Signaling
    • Chromatin Binding Proteins
    • DNA / RNA binding

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human Fibrillarin protein (ab99443)
  • Related Products

    • Nucleolar/Nuclear Staining Kit - Green/ Red Fluorescence - Cytopainter (ab139474)
    • Nucleolar Staining Kit - Green Fluorescence - Cytopainter (ab139475)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab5821 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF (10)
Use a concentration of 0.1 µg/ml.
WB (7)
Use a concentration of 1 µg/ml. Detects a band of approximately 39 kDa (predicted molecular weight: 34 kDa).
Notes
ICC/IF
Use a concentration of 0.1 µg/ml.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 39 kDa (predicted molecular weight: 34 kDa).

Target

  • Function

    S-adenosyl-L-methionine-dependent methyltransferase that has the ability to methylate both RNAs and proteins. Involved in pre-rRNA processing by catalyzing 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. 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).
  • Sequence similarities

    Belongs to the methyltransferase superfamily. Fibrillarin family.
  • Post-translational
    modifications

    By homology to other fibrillarins, some or all of the N-terminal domain arginines are modified to asymmetric dimethylarginine (DMA).
  • Cellular localization

    Nucleus, 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 Drosophila melanogaster
    • Entrez Gene: 2091 Human
    • Entrez Gene: 102643269 Mouse
    • Entrez Gene: 14113 Mouse
    • Omim: 134795 Human
    • SwissProt: Q9W1V3 Drosophila melanogaster
    • SwissProt: P22087 Human
    • SwissProt: P35550 Mouse
    • SwissProt: P22232 Xenopus laevis
    • Unigene: 2821 Drosophila melanogaster
    • Unigene: 299002 Human
    • Unigene: 4595 Mouse
    • Unigene: 8139 Xenopus laevis
    see all
  • Alternative names

    • 34 kD nucleolar scleroderma antigen antibody
    • 34 kDa nucleolar scleroderma antigen antibody
    • Fbl antibody
    • FBRL_HUMAN antibody
    • FIB antibody
    • FIB1 antibody
    • FLRN antibody
    • Histone-glutamine methyltransferase antibody
    • Nop1p antibody
    • RNA U3 small nucleolar interacting protein 1 antibody
    • RNU3IP1 antibody
    • rRNA 2' O methyltransferase fibrillarin antibody
    • rRNA 2'-O-methyltransferase fibrillarin antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody - Nucleolar Marker (ab5821)
    Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody - Nucleolar Marker (ab5821)

    ab5821 staining Fibrillarin in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab5821 at 0.1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

  • Western blot - Anti-Fibrillarin antibody - Nucleolar Marker (ab5821)
    Western blot - Anti-Fibrillarin antibody - Nucleolar Marker (ab5821)
    All lanes : Anti-Fibrillarin antibody - Nucleolar Marker (ab5821) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : Schneider L2 whole cell lysate (ab14893)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size: 34 kDa
    Observed band size: 37 kDa why is the actual band size different from the predicted?
    Additional bands at: 42 kDa, 70 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 3 minutes
  • Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody - Nucleolar Marker (ab5821)
    Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody - Nucleolar Marker (ab5821)This image is courtesy of Luke Hughes-Davies and Rhiannon Jade, Gurdon Institute, Cambridge, UK

    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.

Protocols

  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (158)

Publishing research using ab5821? Please let us know so that we can cite the reference in this datasheet.

ab5821 has been referenced in 158 publications.

  • Giannios P & Casanova J Systemic and local effect of the Drosophila headcase gene and its role in stress protection of Adult Progenitor Cells. PLoS Genet 17:e1009362 (2021). PubMed: 33556132
  • Gaspar VP  et al. Interactome Analysis of KIN (Kin17) Shows New Functions of This Protein. Curr Issues Mol Biol 43:767-781 (2021). PubMed: 34449532
  • Zhang L  et al. lncRNA Sensing of a Viral Suppressor of RNAi Activates Non-canonical Innate Immune Signaling in Drosophila. Cell Host Microbe 27:115-128.e8 (2020). PubMed: 31917956
  • Greig JA  et al. Arginine-Enriched Mixed-Charge Domains Provide Cohesion for Nuclear Speckle Condensation. Mol Cell 77:1237-1250.e4 (2020). PubMed: 32048997
  • Bizhanova A  et al. Distinct features of nucleolus-associated domains in mouse embryonic stem cells. Chromosoma 129:121-139 (2020). PubMed: 32219510
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-10 of 28 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-Fibrillarin antibody - Nucleolar Marker

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Bone Osteosarcoma Epithelial Cells)
Permeabilization
Yes - 0.2% triton
Specification
Bone Osteosarcoma Epithelial Cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Apr 29 2021

Immunocytochemistry/ Immunofluorescence abreview for Anti-Fibrillarin antibody - Nucleolar Marker

Inconclusive
Abreviews
Abreviews
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rhynchosciara americana Cell (salivary gland)
Permeabilization
No
Specification
salivary gland
Fixative
Formaldehyde
Read More

DR. Eduardo Gorab

Verified customer

Submitted Jul 02 2018

Western blot abreview for Anti-Fibrillarin antibody - Nucleolar Marker

Excellent
Abreviews
Abreviews
Application
Western blot
Sample
Mouse Tissue lysate - whole (Whole brain tissue lysate)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (3-8% Tris-Acetate)
Loading amount
15 µg
Specification
Whole brain tissue lysate
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Read More

Abcam user community

Verified customer

Submitted Apr 10 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-Fibrillarin antibody - Nucleolar Marker

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Monkey Cell (Kidney)
Permeabilization
Yes - 0.1% triton x-100
Specification
Kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Jun 10 2016

Western blot abreview for Anti-Fibrillarin antibody - Nucleolar Marker

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Monkey Cell lysate - whole cell (Kidney)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
30 µg
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted May 04 2016

Immunohistochemistry (Frozen sections) abreview for Anti-Fibrillarin antibody - Nucleolar Marker

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
Yes - 0.3% Triton x 100: 30 min
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted May 04 2016

Immunocytochemistry/ Immunofluorescence abreview for Anti-Fibrillarin antibody - Nucleolar Marker

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (mouse embryonic stem cells (E14 mES))
Permeabilization
Yes - 0.25% triton
Specification
mouse embryonic stem cells (E14 mES)
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 0.5% · Temperature: RT°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Jan 05 2016

Western blot abreview for Anti-Fibrillarin antibody - Nucleolar Marker

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - other (Brain)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
20 µg
Specification
Brain
Blocking step
Odyssey Blocking Buffer (PBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 45% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Sep 18 2015

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Fibrillarin antibody - Nucleolar Marker

Good
Abreviews
Abreviews
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
None
Permeabilization
Yes - Tween-20
Specification
Brain
Blocking step
Heat-Inactivated Normal Donkey Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Apr 27 2015

Western blot abreview for Anti-Fibrillarin antibody - Nucleolar Marker

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing
Sample
Xenopus laevis Cell lysate - nuclear (adult heart)
Specification
adult heart
Blocking step
Milk as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
Read More

Abcam user community

Verified customer

Submitted Aug 20 2014

1-10 of 28 Abreviews or Q&A

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