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
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Mouse, Human, Drosophila melanogaster
Predicted to work with: Xenopus laevis
Does not react with:
Rat -
Immunogen
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Positive control
- ICC/IF: HeLa cells
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 < 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
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
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 | 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
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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)Labab5821 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
References (130)
ab5821 has been referenced in 130 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
- García-Aguirre I et al. Enhanced nuclear protein export in premature aging and rescue of the progeria phenotype by modulation of CRM1 activity. Aging Cell 18:e13002 (2019). PubMed: 31305018
- Luczak MW et al. p53 activation by Cr(VI): a transcriptionally limited response induced by ATR kinase in S-phase. Toxicol Sci N/A:N/A (2019). PubMed: 31388677
- Azkanaz M et al. Protein quality control in the nucleolus safeguards recovery of epigenetic regulators after heat shock. Elife 8:N/A (2019). PubMed: 31199242
