Anti-Fibrillarin抗体- Nucleolar Marker
参阅全部 Fibrillarin 一抗
兔多克隆抗体to Fibrillarin - Nucleolar Marker
Rabbit
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.
适用于: ICC/IF, WBmore details
与反应: Mouse, Human, Drosophila melanogaster
预测可用于: Xenopus laevis
不与反应: Rat
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
ICC/IF: HeLa cells
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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Liquid
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.
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
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.
浓度
100 µg 浓度为 1 mg/ml
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab5821于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| ICC/IF | (10) | 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). |
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
SwissProt: P22232 Xenopus laevis
Unigene: 2821 Drosophila melanogaster
Unigene: 299002 Human
Unigene: 4595 Mouse
Unigene: 8139 Xenopus laevis
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RNA U3 small nucleolar interacting protein 1 antibody
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Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody - Nucleolar Marker (ab5821)
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).
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 kDawhy 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.
