Anti-Fibrillarin 抗体 [38F3] - Nucleolar Marker (ab4566)
![Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)](https://www.abcam.co.jp/ps/products/4/ab4566/Images/ab4566_1.jpg "Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)")
Key features and details
- Mouse monoclonal [38F3] to Fibrillarin - Nucleolar Marker
- Suitable for: Flow Cyt, ICC/IF, WB
- Reacts with: Mouse, Rat, Human
- Isotype: IgG1
製品の概要
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製品名
Anti-Fibrillarin antibody [38F3] - Nucleolar Marker
Fibrillarin 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [38F3] to Fibrillarin - Nucleolar Marker -
由来種
Mouse -
特異性
This clone was selected because it stains a single ~34kDa band on western blotting and shows a clear and strong punctate staining of yeast nuclei. It can therefore be used to identify nucleoli immunocytochemically. ab4566 was raised against yeast nuclear preps and the immunogen was identified as Nop1p, the yeast homolog of fibrillarin. Due to high aa homology the antibody should work with any specie possessing a nucleus, however this has not been tested.
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アプリケーション
適用あり: Flow Cyt, ICC/IF, WBmore details -
種交差性
交差種: Mouse, Rat, Human
交差が予測される動物種: Plants
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免疫原
Tissue, cells or virus corresponding to Saccharomyces cerevisiae Fibrillarin.
Yeast nuclear preparation (S. cerevisiae). Hybridomas were screened by immunofluorescence on yeast cells and by western blotting on yeast protein homogenates (S. cerevisiae). -
ポジティブ･コントロール
- ICC: HEK-293 cells and SH-SY5Y cells; WB: HEK-293, C6, NIH-3T3 nuclear fractions
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特記事項
Pfam number: PF01269. A reference below describes the characterization of D77, an antibody very similar but not identical to ab4566.
Gives a much weaker signal in western blot compared to ab218846.
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.
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製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
バッファー
Preservative: 0.065% Sodium azide
Constituent: Tissue culture supernatant -
Concentration information loading... -
精製度
Tissue culture supernatant -
特記事項（精製）
Sterile filtered. -
ポリ/モノ
モノクローナル -
クローン名
38F3 -
アイソタイプ
IgG1 -
研究分野
関連製品
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Compatible Secondaries
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Isotype control
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Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab4566の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
| アプリケーション | Abreviews | 特記事項 |
|---|---|---|
| Flow Cyt |
1/20.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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| ICC/IF | (8) |
1/100.
For IF of mammalian cells 1/500. ab4566 is sensitive to aldehyde. Use a mild formalin fixation or acetone or methanol fixation as the target is nuclear. |
| WB | (10) |
1/2000 - 1/10000. Detects a band of approximately 34 kDa. 1/2000 (cell lysates) - 1/10000 (nuclear fractions)(ECL). For other (non-ECL) western detection methods, 1/1000 - 1/5000. To detect mammalian fibrillarin on western blots by ECL, 1/500.
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| 特記事項 |
|---|
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Flow Cyt
1/20. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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ICC/IF
1/100. For IF of mammalian cells 1/500. ab4566 is sensitive to aldehyde. Use a mild formalin fixation or acetone or methanol fixation as the target is nuclear. |
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WB
1/2000 - 1/10000. Detects a band of approximately 34 kDa. 1/2000 (cell lysates) - 1/10000 (nuclear fractions)(ECL). For other (non-ECL) western detection methods, 1/1000 - 1/5000. To detect mammalian fibrillarin on western blots by ECL, 1/500. |
ターゲット情報
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機能
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). -
配列類似性
Belongs to the methyltransferase superfamily. Fibrillarin family. -
翻訳後修飾
By homology to other fibrillarins, some or all of the N-terminal domain arginines are modified to asymmetric dimethylarginine (DMA). -
細胞内局在
Nucleus, nucleolus. Fibrillar region of the nucleolus. - Information by UniProt
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参照データベース
- Entrez Gene: 2091 Human
- Entrez Gene: 102643269 Mouse
- Entrez Gene: 14113 Mouse
- Entrez Gene: 292747 Rat
- Omim: 134795 Human
- SwissProt: P22087 Human
- SwissProt: P35550 Mouse
- SwissProt: P22509 Rat
see all -
別名
- 34 kD nucleolar scleroderma antigen antibody
- 34 kDa nucleolar scleroderma antigen antibody
- Fbl antibody
see all
画像
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Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)
Rat neurons and glial stained with mouse monoclonal to Fibrillarin (green) and with chicken antibody to neurofilament NF-H (red). Cells were counterstained with a fluorescent DNA probe (blue). Nuclear DNA is revealed with Hoechst dye (blue). Cultures were processed using our standard fixation and staining procedure (in protocol section).
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![Immunocytochemistry - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)](https://www.abcam.co.jp/ps/products/4/ab4566/Images/ab4566-342869-AntiFibrillarinantibody38F3helahuman.jpg)
Immunocytochemistry - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)ICC analysis of HeLa cells stained with mouse monoclonal to Fibrillarin (green) and with chicken antibody to vimentin (red) and counterstained with a fluorescent DNA probe (blue). Nuclear DNA is revealed with DAPI(blue). The vimentin antibody was used at a dilution of 1/1000 and the fibrillarin monoclonal at 1/100. Cultures were processed using standard fixation and staining procedure (in protocol section).
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![Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)](https://www.abcam.co.jp/ps/products/4/ab4566/Images/ab4566-342868-AntiFibrillarinantibody38F3westernblot.jpg)
Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)All lanes : Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566) at 1/500 dilution
Lane 2 : C6 cytosol fraction
Lane 3 : C6 nuclear fraction
Lane 4 : HEK-293 cytosol fraction
Lane 5 : HEK-293 nuclear fraction
Lane 6 : NIH-3T3 cytosol fraction
Lane 7 : NIH-3T3 nuclear fraction
Observed band size: 37 kDa why is the actual band size different from the predicted? -
![Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)](https://www.abcam.co.jp/ps/products/4/ab4566/Images/ab4566_2.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)Human neuroblastoma line SH-SY5Y stained with mouse monoclonal to Fibrillarin (green) and with chicken antibody to neurofilament NF-H (red) and counterstained with a fluorescent DNA probe (blue). Nuclear DNA is revealed with Hoechst dye (blue). The NF-H antibody was used at a dilution of 1/100000 and the fibrillarin monoclonal at 1/1000. Cultures were processed using standard fixation and staining procedure (in protocol section).
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![Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)](https://www.abcam.co.jp/ps/products/4/ab4566/Images/ab4566-9284-anti-fibrillarin-antibody-38f3-western-blot.jpg)
Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)Image courtesy of an anonymous Abreview.Mouse embryonic fibroblast fractionation.
Cytopl - cytoplasmic fraction.Nucl - nuclear fraction.
20 µg of each loaded.
ab4566 used at a 1/2000 dilution.The secondary used was an Alexa-Fluor 680 conjugated goat anti-mouse polyclonal used at a 1/10000 dilution. -
![Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)](https://www.abcam.co.jp/ps/products/4/ab4566/Images/ab4566-262968-anti-fibrillarin-antibody-38f3-western-blot.jpg)
Western blot - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)Image is courtesy of Dr Svetlana KhoronenkovaAll lanes : Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566) at 1/2000 dilution (incubated for 1 hour, diluted with Blocking buffer 1:1 in PBS+0.1%Tween)
Lane 1 : cytoplasmic protein fraction of HeLa cells with LI-COR® Odyssey® Blocking Buffer, 45 minutes at room temperature at 50 %
Lane 2 : nuclear protein fraction of HeLa cells with LI-COR® Odyssey® Blocking Buffer, 45 minutes at room temperature at 50 %
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : AlexaFluor 680 goat anti-mouse at 1/10000 dilution
Performed under reducing conditions.
Additional bands at: 34 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 second
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![Flow Cytometry - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)](https://www.abcam.co.jp/ps/products/4/ab4566/Images/ab4566-9285-anti-fibrillarin-antibody-38f3-flow-cytometry.jpg)
Flow Cytometry - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)Overlay histogram showing HEK293 cells stained with ab4566 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab4566, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1](ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. -
![Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)](https://www.abcam.co.jp/ps/products/4/ab4566/Images/ab4566-262967-anti-fibrillarin-antibody-38f3-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)Image is courtesy of Cesar Camachoab staining Fibrillarin in Human melanoma A7 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton and blocked with 1% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (1/500 in 1% BSA) for 24 hours at 4°C. A FITC-conjugated Goat anti-mouse polyclonal (1/200) was used as the secondary antibody.
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![Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)](https://www.abcam.co.jp/ps/products/4/ab4566/Images/ab4566_3.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Fibrillarin antibody [38F3] - Nucleolar Marker (ab4566)High magnification view of human Hek293 cell nuclei stained with mouse monoclonal to fibrillarin (green), counterstained with a fluorescent DNA probe (blue). Nuclear DNA is revealed with Hoechst dye (blue). Cultures were processed using our standard fixation and staining procedure (in protocol section).
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (175)
ab4566 は 175 報の論文で使用されています。
- Kim ES et al. C9orf72-associated arginine-rich dipeptide repeats induce RNA-dependent nuclear accumulation of Staufen in neurons. Hum Mol Genet 30:1084-1100 (2021). PubMed: 33783499
- Villacampa A et al. From Spaceflight to Mars g-Levels: Adaptive Response of A. Thaliana Seedlings in a Reduced Gravity Environment Is Enhanced by Red-Light Photostimulation. Int J Mol Sci 22:N/A (2021). PubMed: 33477454
- Hasygar K et al. Coordinated control of adiposity and growth by anti-anabolic kinase ERK7. EMBO Rep 22:e49602 (2021). PubMed: 33369866
- Weeks SE et al. Inhibiting ß-catenin disables nucleolar functions in triple-negative breast cancer. Cell Death Dis 12:242 (2021). PubMed: 33664239
- Zhu Q et al. Rcl1 depletion impairs 18S pre-rRNA processing at the A1-site and up-regulates a cohort of ribosome biogenesis genes in zebrafish. Nucleic Acids Res 49:5743-5759 (2021). PubMed: 34019640