Anti-Fibrillarin antibody - Nucleolar Marker (ab5821)
- Product nameAnti-Fibrillarin antibody - Nucleolar MarkerSee 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 applicationsIHC - Wholemount, ICC/IF, IP, IHC-P, WB, ICC more details
- Species reactivityReacts with: Mouse, Human, Xenopus laevis, Fruit fly (Drosophila melanogaster)
Does not react withRat
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Fibrillarin.
(Peptide available as ab13745.)
- Positive control
- This antibody gave a positive signal in the following whole cell lysates: HeLa; A431; Jurkat; 293T. This antibody also gave a positive signal in HeLa nuclear lysate.
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
- Concentration information loading...
- PurityImmunogen affinity purified
- Clonality Polyclonal
- Research Areas
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.
|IHC - Wholemount||IHC - Wmt: Use at an assay dependent concentration. PubMed: 22291607|
|ICC/IF||ICC/IF: Use at an assay dependent dilution.|
|IP||IP: Use at an assay dependent dilution. PubMed: 19430468|
|WB||WB: Use a concentration of 1 µg/ml. Detects a band of approximately 39 kDa (predicted molecular weight: 34 kDa).Can be blocked with Human Fibrillarin peptide (ab13745).|
- FunctionInvolved in pre-rRNA processing. Utilizes the methyl donor S-adenosyl-L-methionine to catalyze 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.
- 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.
- Entrez Gene: 37662 Fruit fly (Drosophila melanogaster)
- Entrez Gene: 2091 Human
- Entrez Gene: 100044829 Mouse
- Entrez Gene: 14113 Mouse
- Omim: 134795 Human
- SwissProt: Q9W1V3 Fruit fly (Drosophila melanogaster)
- SwissProt: P22087 Human
- SwissProt: P35550 Mouse
- SwissProt: P22232 Xenopus laevis
- Unigene: 2821 Fruit fly (Drosophila melanogaster)
- Unigene: 299002 Human
- Unigene: 4595 Mouse
- Unigene: 8139 Xenopus laevis
- 34 kD nucleolar scleroderma antigen antibody
- 34 kDa nucleolar scleroderma antigen antibody
- 34-kD nucleolar scleroderma antigen antibody
- FBL antibody
- FBRL_HUMAN antibody
- FIB antibody
- FIB1 antibody
- FLRN 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
Anti-Fibrillarin antibody - Nucleolar Marker images
The cells were 100% methanol fixed (5 min) and then
incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab37266, 1µg/ml, red) and (ab5821, 1µg/ml, blue) overnight at +4°C. The secondary antibodies were ab150115 Alexa Fluor® 647 goat anti-mouse IgG (H+L) used at 2µg/ml for 1h and ab175652 Alexa Fluor® 405 goat anti-rabbit IgG (H+L) used at 2µg/ml for 1h.
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).
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 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.
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 (Fruit fly (Drosophila melanogaster)) Whole Cell Lysate (ab14893)
Lysates/proteins at 20 µg per lane.
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
Immunohistochemical analysis of salivary glands from Drosophila melanogaster larvae, staining Fibrillarin with ab5821. Tissues were fixed in 4% paraformaldehyde/PBS and 0.3% Triton-X/PBS before incubation with primary antibody.
References for Anti-Fibrillarin antibody - Nucleolar Marker (ab5821)
This product has been referenced in:
- Schenkwein D et al. rDNA-directed integration by an HIV-1 integrase--I-PpoI fusion protein. Nucleic Acids Res 41:e61 (2013). ICC/IF ; Human . Read more (PubMed: 23275537) »
- Madlener S et al. Essential role for mammalian apurinic/apyrimidinic (AP) endonuclease Ape1/Ref-1 in telomere maintenance. Proc Natl Acad Sci U S A 110:17844-9 (2013). Read more (PubMed: 24127576) »