Anti-Fibrillarin antibody ab5821 is a rabbit polyclonal antibody that is used in Fibrillarin western blotting and immunofluorescence. Suitable for human and mouse samples.
- Tried and trusted by researchers since 2004
IgG
Rabbit
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Liquid
Polyclonal
WB | ICC/IF | |
---|---|---|
Human | Tested | Tested |
Mouse | Tested | Expected |
Rat | Not recommended | Not recommended |
Drosophila melanogaster | Tested | Expected |
Xenopus laevis | Not recommended | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1 µg/mL | Notes - |
Species Drosophila melanogaster | Dilution info 1 µg/mL | Notes - |
Species Human | Dilution info 1 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Xenopus laevis | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 0.1 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Drosophila melanogaster | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Xenopus laevis | Dilution info - | Notes - |
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S-adenosyl-L-methionine-dependent methyltransferase that has the ability to methylate both RNAs and proteins (PubMed:24352239, PubMed:30540930, PubMed:32017898). Involved in pre-rRNA processing by catalyzing the site-specific 2'-hydroxyl methylation of ribose moieties in pre-ribosomal RNA (PubMed:30540930). Site specificity is provided by a guide RNA that base pairs with the substrate (By similarity). Methylation occurs at a characteristic distance from the sequence involved in base pairing with the guide RNA (By similarity). Probably catalyzes 2'-O-methylation of U6 snRNAs in box C/D RNP complexes (PubMed:32017898). U6 snRNA 2'-O-methylation is required for mRNA splicing fidelity (PubMed:32017898). 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, PubMed:30540930). Part of the small subunit (SSU) processome, first precursor of the small eukaryotic ribosomal subunit. During the assembly of the SSU processome in the nucleolus, many ribosome biogenesis factors, an RNA chaperone and ribosomal proteins associate with the nascent pre-rRNA and work in concert to generate RNA folding, modifications, rearrangements and cleavage as well as targeted degradation of pre-ribosomal RNA by the RNA exosome (PubMed:34516797).
FIB1, FLRN, FBL, rRNA 2'-O-methyltransferase fibrillarin, 34 kDa nucleolar scleroderma antigen, Histone-glutamine methyltransferase, U6 snRNA 2'-O-methyltransferase fibrillarin
Anti-Fibrillarin antibody ab5821 is a rabbit polyclonal antibody that is used in Fibrillarin western blotting and immunofluorescence. Suitable for human and mouse samples.
- Tried and trusted by researchers since 2004
IgG
Rabbit
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Liquid
Polyclonal
Affinity purification Immunogen
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.
Replenishment batches of our polyclonal antibody, ab5821 are tested in WB. Previous batches were additionally validated in ICC/IF. This application is still expected to work and is covered by our Abpromise guarantee. You may also be interested in our alternative recombinant antibody, ab166630.
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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Full details and terms and conditions can be found here:
Terms & Conditions.
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 Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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: Western blot - Anti-Fibrillarin antibody - Nucleolar Marker (ab5821) at 1 µg/mL
Lane 1: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg
Lane 2: NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 20 µg
Lane 3: Schneider L2 whole cell lysate (ab14893) at 20 µg
All lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 33 kDa
Observed band size: 37 kDa, 42 kDa, 70 kDa
Exposure time: 3min
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.
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