Key features and details
- Rabbit polyclonal to Fibrillarin - Nucleolar Marker
- Suitable for: ICC/IF, WB
- Reacts with: Mouse, Human, Drosophila melanogaster
- Isotype: IgG
Product nameAnti-Fibrillarin antibody - Nucleolar Marker
See all Fibrillarin primary antibodies
DescriptionRabbit polyclonal to Fibrillarin - Nucleolar Marker
SpecificityThis 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 applicationsSuitable for: ICC/IF, WBmore details
Species reactivityReacts with: Mouse, Human, Drosophila melanogaster
Predicted to work with: Xenopus laevisDoes not react with: Rat
- ICC/IF: HeLa cells
Storage instructionsShipped 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 bufferpH: 7.40
Preservative: 0.02% Sodium azide
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...
PurityImmunogen affinity purified
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.
|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).|
FunctionS-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 similaritiesBelongs to the methyltransferase superfamily. Fibrillarin family.
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.
- Information by UniProt
- 34 kD nucleolar scleroderma antigen antibody
- 34 kDa nucleolar scleroderma antigen antibody
- Fbl antibody
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).
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.
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
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.
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