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
Overview
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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 & Species
See all applications and species dataApplication Species ICC/IF HumanWB Human
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- ICC/IF: HeLa cells
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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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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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
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
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.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanWB HumanAll applications Xenopus laevisApplication Abreviews Notes ICC/IF (9) 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
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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
modificationsBy 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. - Information by UniProt
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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
see all -
Alternative names
- 34 kD nucleolar scleroderma antigen antibody
- 34 kDa nucleolar scleroderma antigen antibody
- Fbl antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody - Nucleolar Marker (ab5821)Lab
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).
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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)This image is courtesy of Luke Hughes-Davies and Rhiannon Jade, Gurdon Institute, Cambridge, UKImmunofluorescent 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
Datasheets and documents
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SDS download
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Datasheet download
References (140)
ab5821 has been referenced in 140 publications.
- Martínez Corrales G et al. Partial Inhibition of RNA Polymerase I Promotes Animal Health and Longevity. Cell Rep 30:1661-1669.e4 (2020). PubMed: 32049000
- Gershman BW et al. Tissue-specific Expression of Ribosomal Protein Paralogue eRpL22-like in Drosophila melanogaster Eye Development. Dev Dyn N/A:N/A (2020). PubMed: 32353187
- Watson SF et al. ILF3 contributes to the establishment of the antiviral type I interferon program. Nucleic Acids Res 48:116-129 (2020). PubMed: 31701124
- Alexandrova J et al. Full-length NF-?B repressing factor contains an XRN2 binding domain. Biochem J 477:773-786 (2020). PubMed: 32011671
- Chen L et al. Loss of Human TGS1 Hypermethylase Promotes Increased Telomerase RNA and Telomere Elongation. Cell Rep 30:1358-1372.e5 (2020). PubMed: 32023455
Images
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Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody - Nucleolar Marker (ab5821) Lab
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).
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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) 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.
