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Fibrillarin

Fibrillarin is a nucleolar protein involved in the processing of pre-ribosomal RNA and the methylation of ribosomal RNA.
It plays a crucial role in ribosome biogenesis and is essential for cell viability.
Fibrillarin is a highly conserved protein found in eukaryotes and archaea, and its dysfunction has been implicated in various disease states, including cancer and autoimmune disorders.
Researching Fibrillarin is essential for understanding fundamental cellular processes and developing potential therapeutic interventions.
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Most cited protocols related to «Fibrillarin»

1
Constructing mNeonGreen Fusion Protein Vectors
Cited 67 times
mNeonGreen fluorescent protein expression vectors were constructed using C1 and N1 (Clontech-style) cloning vectors. The mNeonGreen cDNA was amplified with a 5′ primer encoding an AgeI site and a 3′ primer encoding either a BspEI (C1) or NotI (N1) site for generating cloning vectors to create C-terminal and N-terminal fusions (with regards to the FP), respectively. Purified and digested PCR products were ligated into similarly digested EGFP-C1 and EGFP-N1 cloning vector backbones. To obtain targeting fusion vectors, the appropriate cloning vector and a previously assembled EGFP or mEmerald fusion vector were digested, either sequentially or doubly, with the appropriate enzymes and ligated together after gel purification.
Thus, to prepare mNeonGreen C-terminal fusions (number of linker amino acids in parenthesis), the following digests were performed: annexin A4 (12), NheI and BspEI (Alen Piljic, EMBL, Heidelberg, Germany; NM_001153.3); β-actin (7), NheI and BglII (human β-actin cDNA source: Clontech; NM_001101.3); β-catenin (20), XhoI and BamHI (mouse β-catenin cDNA source: Origene, Rockville, MD; NM_001165902.1); 20 amino acid farnesylation signal from c-Ha-Ras (CAAX; 5), AgeI and BspEI (c-Ha-Ras cDNA source: Clontech; NM_001130442.1); CAF1 (10), AgeI and BspEI (mouse chromatin assembly factor cDNA source: Akash Gunjan, Florida State University; NM_013733.3); caveolin 1 (10), NheI and BglII (human caveolin 1 cDNA source: Origene; NM_001753); endosomes (14), NheI and BspEI (endosomes cDNA source: Clontech; NM_004040.2); fascin (10), BspEI and BamHI (human fascin cDNA source: Origene; NM_003088.2); fibrillarin (7), AgeI and BspEI (fibrillarin cDNA source: Evrogen, Moscow, Russia; NM_001436.3); filamin A (14), BspEI and HindIII (human filamin cDNA source: David Calderwood, Yale University; NM_001456.3); human lysosomal membrane glycoprotein 1 (20), BamHI and NotI (LAMP1; George Patterson, NIH, Bethesda MD, U.S.A.; NM_012857.1); human light chain clathrin (15), NheI and BglII (human clathrin light chain cDNA source: George Patterson, NIH; NM_001834.2); human myotilin, AgeI and BspEI (MYOT; Origene; NM_006790.1); PCNA (19), AgeI and BspEI (proliferating cell nuclear antigen cDNA source: David Gilbert, FSU; NM_002592.2); plastin (10), BspEI and XhoI (human plastin 1 (fimbrin) cDNA source: Origene; NM_002670.1); canine Rab4a, BglII and BamHI (Rab4a cDNA source: Viki Allen, U. Manchester, UK; NM_004578.2); LC3B (7), AgeI and BspEI (rat LC3B cDNA source: Jenny M. Tam, Harvard University; U05784.1); talin (22) AgeI and BspEI (mouse talin 1 cDNA source: Clare Waterman, NIH; NM_011602.5); α-tubulin (18), NheI and BglII (human α-tubulin cDNA source: Clontech; NM_006082).
To prepare mNeonGreen N-terminal fusions (number of linker amino acids in parenthesis), the following digests were performed: human non-muscle α-actinin, EcoRI and NotI (cDNA source, Tom Keller, Florida State University (FSU), Tallahassee, FL, U.S.A.; NM_001130005.1); human calnexin, AgeI and NotI (Origene; NM_001746.3); c-src (7), BamHI and EcoRI (chicken c-src cDNA source: Marilyn Resh, Sloan-Kettering, New York; XM_001232484.1); connexin-43 (7), BamHI and NotI (rat C×43 cDNA source: Matthias Falk, Lehigh U; NM_001004099.1); EB3 (7), BglII and BamHI (EB3 cDNA source: Lynne Cassimeris, Lehigh University; NM_012326.2); human keratin 18, EcoRI and NotI (Open Biosystems; NM_199187.1); lamin B1 (10), EcoRI and BamHI (human lamin B1 cDNA source: George Patterson, NIH; NM_005573.2); Lifeact (7), BamHI and NotI (Lifeact cDNA source: IDT); mouse mannosidase 2 (112 N-terminal amino acids, MANNII; 10), NheI and BamHI (cDNA source: Jennifer Lippincott-Schwartz, NIH; NM_008549.2); myosin IIA (14) NheI and BglII (mouse myosin IIA cDNA source: Origene; NM_022410.2); human nucleoporin 50kDa, BamHI and NotI (NUP50 cDNA source: Origene; NM_007172.2); human pyruvate dehydrogenase, AgeI and NotI (human PDHA1 cDNA source: Origene; NM_000284); human peroxisomal membrane protein, NotI and AgeI (PMP cDNA source: Origene; NM_018663.1); human MAP Tau (10), AgeI and NotI (MAP Tau cDNA source: Origene; NM_016841); human TfR (20), BamHI and NotI (transferrin receptor cDNA source: George Patterson, NIH; NM_NM_003234); human TPX2 (10), AgeI and NotI (TPX2 cDNA source: Patricia Wadsworth, University of Massachusetts, Amherst; NM_012112.4); mouse VASP (10), NheI and BamHI (cDNA source: Clare Waterman, NIH; NM_009499); vascular epithelial cadherin (10), AgeI and NotI (human VE cadherin cDNA source: Origene; NM_001795.3), vimentin (7), BamHI and NotI (human vimentin cDNA source: Robert Goldman, Northwestern University; NM_003380.3), zyxin (6), BamHI and NotI (human zyxin cDNA source: Origene; NM_003461). All DNA for transfection was prepared using the Plasmid Maxi kit (QIAGEN). To ensure proper localization, mNeonGreen fusion proteins were characterized by transfection in HeLa (S3 or CCL2 line) or MDCK cells (ATCC) using Effectene (QIAGEN) and ∼1 μg vector. Transfected cells were grown on coverslips in DMEM/F12, fixed after 48 hours, and mounted with Gelvatol. Epifluorescence images were captured with a Nikon 80i microscope using widefield illumination and a Chroma FITC filter set to confirm proper localization.
Shaner N.C., Lambert G.G., Chammas A., Ni Y., Cranfill P.J., Baird M.A., Sell B.R., Allen J.R., Day R.N., Israelsson M., Davidson M.W, & Wang J. (2013). A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum. Nature methods, 10(5), 407-409.
Publication 2013
2
Construction and Characterization of mTagBFP2 Fusion Proteins
Cited 22 times
mTagBFP2 fluorescent protein expression vectors were constructed using -C1 and -N1 (Clontech-style) cloning vectors. The mTagBFP2 cDNA was amplified with a 5′ primer encoding an AgeI site and a 3′ primer encoding either a BspEI (-C1) or NotI (-N1) site for C-terminal and N-terminal fusions (with regards to the FP), respectively. Purified and digested PCR products were ligated into similarly digested pEGFP-C1 and pEGFP-N1 cloning vector backbones. To generate targeting fusion vectors, the appropriate cloning vector and a previously assembled EGFP fusion vector were digested, either sequentially or doubly, with the appropriate enzymes and ligated together after gel purification.
Thus, to prepare mTagBFP2 C-terminal fusions (number of linker amino acids in parenthesis), the following digests were performed: human lamin B1 (10), NheI and BglII (lamin B1 cDNA source: George Patterson, NIH; NM_005573.2); 20 amino acid farnesylation signal from c-Ha-Ras (CAAX; 5), AgeI and BspEI (c-Ha-Ras cDNA source: Clontech, Mountain View, CA; NM_001130442.1); endoplasmic reticulum (5), AgeI and BspEI (calreticulin cDNA source: George Patterson, NIH; NM_004343.3); fibrillarin (7), AgeI and BspEI (fibrillarin cDNA source: Evrogen, Moscow, Russia; NM_001436.3); human light chain clathrin (15), NheI and BglII (clathrin cDNA source: George Patterson, NIH; NM_001834.2); β-actin (7), NheI and BglII (human β-actin cDNA source: Clontech, Mountain View, CA; NM_001101.3); caveolin 1 (10), NheI and BglII (human caveolin 1 cDNA source: Origene, Rockville, MD; NM_001753); vinculin (22) AgeI and EcoRI (human vinculin source: Clare Waterman, NIH; NM_003373.3); CAF1 (10), AgeI and BspEI (mouse chromatin assembly factor cDNA source: Akash Gunjan, FSU; NM_013733.3) Rab5a (7), NheI and BglII (canine Rab5a cDNA source: Vicki Allen, University of Manchester; NM_001003317.1); α-tubulin (18), NheI and BglII (human α-tubulin cDNA source: Clontech, Mountain View, CA; NM_006082); myosin IIA (18) NheI and BglII (human myosin heavy chain IIA cDNA source: DNA2.0, Menlo Park, CA; AJ312390.1); PCNA (19), AgeI and BspEI (proliferating cell nuclear antigen cDNA source: David Gilbert, FSU; NM_002592.2).
To prepare mTagBFP2 N-terminal fusions (number of linker amino acids in parenthesis), the following digests were performed: β-2 connexin-26 (7), BamHI and NotI (rat Cx26 cDNA source: Matthias Falk, Lehigh U; NM_001004099.1); TfR (20), BamHI and NotI (transferrin receptor cDNA source: George Patterson, NIH; NM_NM_003234); Golgi complex (7), BamHI and NotI (human β-galactosamide α-2,6-sialyltransferase 1cDNA source: Jennifer Lippincott-Schwartz, NIH; NM_173216.2); zyxin (6), BamHI and NotI (human zyxin cDNA source: Origene, Rockville, MD; NM_003461); vascular epithelial cadherin (10), AgeI and NotI (human VE cadherin cDNA source: Origene, Rockville, MD; NM_001795.3); mitochondria (7), BamHI and NotI (human mitochondrial targeting sequence, cytochrome c oxidase cDNA source: Clontech, Mountain View, CA; NM_004074.2); centromere protein B (22), BamHI and NotI (human CENPB cDNA source: Alexey Khodjakov, Wadsworth Center, Albany, NY; NM_001810.5); α-actinin (19), BamHI and EcoRI (human α-actinin cDNA source: Tom Keller, Florida State University, Tallahassee; NM_001130005.1); c-src sarcoma (7), BamHI and EcoRI (chicken c-src cDNA source: Marilyn Resh, Sloan-Kettering, New York; XM_001232484.1); Lifeact (7), BamHI and NotI (Lifeact cDNA source: IDT, Coralville, IA); vimentin (7), BamHI and NotI (human vimentin cDNA source: Robert Goldman, Northwestern University; NM_003380.3).
All DNA for transfection was prepared using the Plasmid Maxi kit (QIAGEN, Valencia, CA). To ensure proper localization, mTagBFP2 fusion proteins were characterized by transfection in HeLa cells (CCL2 line; ATCC, Manassas, VA) using Effectene (QIAGEN) and 1 µg vector. Transfected cells were grown on coverslips in DMEM/F12, fixed after 48 hours, and mounted with Gelvatol. Epifluorescence images (Figure 4) were taken with a Nikon 80i microscope using widefield illumination and an Omega QMax Blue filter set to confirm proper localization.
Subach O.M., Cranfill P.J., Davidson M.W, & Verkhusha V.V. (2011). An Enhanced Monomeric Blue Fluorescent Protein with the High Chemical Stability of the Chromophore. PLoS ONE, 6(12), e28674.
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Publication 2011
3
Construction and Characterization of Nuclear Transporters
Cited 8 times
Strains carrying null alleles, GFP or cherry-mRFP C-terminally tagged fusions of nuclear transporters, and nuclear and nucleolar markers were generated as described (Yang et al., 2004 (link); Supplemental Figure S2), using Aspergillus fumigatus pyrG, riboB, or pyroA gene as a prototrophic selection marker. To construct an N-terminal GFP-tagged KapJKap123, a null kapJKap123::pyrGAf strain was transformed with a 5′UTR::GFP::KapJORF::3′UTR DNA cassette, and the replacement of the null locus was selected by growing the transformants in regeneration plates containing 2 mg/ml 5-fluoro-orotic acid (5-FOA; Apollo Scientific, Stockport, United Kingdom). After 24 h of incubation at 37°C, a layer of Aspergillus minimal medium containing the required nutrients and supplements plus 2 mg/ml FOA was poured onto the protoplast-regeneration plates. Colonies that grew through this layer of medium were purified to homokaryosis and analyzed by Southern blot to verify the correct integration of the cassette at the null kapJKap123 locus. Strains coexpressing tagged karyopherins and histone H1 (HhoA::mCh) or fibrillarin (An-Fib::mCh) were obtained by step-by-step transformation of MAD1425 with each fusion PCR cassette. Amplification of HhoA and An-Fib tagging cassettes follow standard procedures as described, but the selectable marker in this case was A. fumigatus pyroA gene. Strains MAD2653 (kapLMsn5::gfp; hhoA::mCh) and MAD2654 (kapIPse1::gfp; hhoA::mCh) were obtained by transformation of MAD2446 with the karyopherin-tagging PCR cassette. Strains MAD3772 (kapHKap120::gfp; nls::Dsred) and MAD3771 (kapIPse1::gfp; nls::Dsred) were obtained by transformation of AY02 with the karyopherin-tagging PCR cassette. Strain MAD3817 is a diploid obtained from MAD3773 and MAD3604 haploid strains.
Oligonucleotides used in this study are summarized in Supplementary Table S2. Homologous recombination for each construct was confirmed by both diagnostic PCR and Southern blot analysis.
The expression of tagged proteins was analyzed by Western blotting using standard procedures. Briefly, A. nidulans strains were cultivated for 18 h in fermentation medium (Orejas et al., 1995 (link)), filtered through Miracloth (Calbiochem, La Jolla, CA), squeezed to dry, frozen in dry ice, and lyophilized for 16 h. Protein extraction was carried out as previously described (Araújo-Bazán et al., 2008 (link)), and 50 μg were loaded on an 8% polyacrylamide gel before electrotransfer to nitrocellulose filters. To detect GFP fusions, filters were incubated with anti-GFP mouse monoclonal antibody cocktail (1/5000; Roche, Indianapolis, IN). Actin, used as loading and extract quality control, was detected using mouse anti-actin antibody (1/50,000; ICN Biomedical, Irvine, CA). Peroxidase-conjugated anti-mouse (1/4000; Jackson ImmunoResearch Laboratories, West Grove, PA) was used as secondary antibody.
To characterize respective transporter coding sequence (CDS), cDNA was generated from total RNA extracted using TRIzol reagent (Invitrogen, Carlsbad, CA) and following the manufacturer's instructions. Total RNA was isolated from mycelia of a wild-type strain cultured under standard conditions. A single-strand cDNA library was made using the First-Strand cDNA Kit (Roche) and an oligo-dT primer. cDNA for each nuclear transporter was amplified using gene-specific oligonucleotides, and the CDS was confirmed by sequencing.
Markina-Iñarrairaegui A., Etxebeste O., Herrero-García E., Araújo-Bazán L., Fernández-Martínez J., Flores J.A., Osmani S.A, & Espeso E.A. (2011). Nuclear transporters in a multinucleated organism: functional and localization analyses in Aspergillus nidulans. Molecular Biology of the Cell, 22(20), 3874-3886.
Publication 2011
4
High-throughput siRNA Screening of Nucleolar Proteins
Cited 7 times

Protocol full text hidden due to copyright restrictions

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Publication 2018
Alexa Fluor 647 Anti-Antibodies cDNA Library Cell Nucleus Cells Fetal Bovine Serum fibrillarin Fluorescent Antibody Technique Goat HOE 33342 Immunoglobulin G Light Lipofectamine Mus paraform RNA, Small Interfering Technique, Dilution Triton X-100
5
Immunostaining of C. elegans Male Gonads
Cited 7 times
Male gonads were dissected in 5–10 microliters of sperm salts on ColorFrost Plus slides (Fisher Scientific) using established protocols for antibody staining of C. elegans gonads provided in Wormbook [92] . Three different fixation methods were used in this study. For paraformaldehyde staining, animals were processed as described in [61] (link),[92] . For cold methanol or methanol/acetone fixation, animals were dissected in sperm salts, and then a coverslip with four corner dots of silicon grease was placed over the isolated gonad and gentle pressure was applied to generate partially flattened gonads and/or monolayers of spermatocytes and spermatids. The slide preparation was then placed either in liquid nitrogen or on dry ice. After freezing, the coverslip was removed. For methanol/acetone preparations, the slide was immersed in 95% methanol for 10 minutes followed by a 5 minute immersion in 100% acetone. Slides were allowed to air dry briefly. For −20°C methanol preparations, slides were kept in methanol overnight. Slides were washed with three consecutive 10 minute washes in PBS followed by a 30 min. room temperature incubation in blocking solution (PBS+0.5% BSA and 0.1% Tween 20). Primary and secondary antibody incubations were each diluted into blocking solution at conducted at room temperature in a humid chamber.
For DIC/Hoechst preparations, Hoechst 33342 (Sigma-Aldrich) was used at 100 µg/ml.
The following primary antibodies were used in overnight incubations (unless otherwise noted) with different fixation conditions. Commercial sources or labs kindly providing antibodies are also listed.
Paraformaldehyde fixed preparations: 1∶200 mouse anti-REC-8 (Abcam); mouse anti-Nop-1 (yeast fibrillarin mAbD77, Aris lab) used at a 1∶1000 dilution [93] (link), rabbit anti-HIM-3 (Zetka Lab) used at a 1∶400 dilution [44] (link), rabbit anti-CeLamin (Gruenbaum lab) used at a 1∶500 dilution, rabbit anti-AIR-2 (Schumacher lab) used at a 1∶500 dilution, rabbit anti-SPD-2 (O'Connell Lab) used at a 1∶500 dilution, guinea pig anti-SYP-1 (Villeneuve Lab) was preasborbed against homozygote syp-1(me17) mutant animals from the strain AV307 and used at a 1∶200 dilution [45] (link).
Methanol-acetone fixed preparations: rabbit anti-HCP-1 used at a 1∶200 dilution , rabbit anti-HCP-3 used at a 1∶200 dilution, and rabbit anti-HCP-4 used at a 1∶200 dilution (Moore lab), rabbit anti-HIM-10 (Meyer lab) used at a 1∶500 dilution [61] (link). The HCP-2 antibody, used at a 1∶200 dilution, is a rabbit polyclonal raised against the peptide NSVDDNSYCEPPRASSAHD that correspond to amino acids 93–110 of HCP-2.
Cold methanol preparations as described in [94]: 1∶400 rabbit anti-pHisH3-ser10 (Upstate Biotechnology), 1∶100 FITC-conjugated anti-α-tubulin (DM1A; Sigma-Aldrich), 1∶1000 rabbit anti-PLK-1 [55] (link) (Golden Lab). 1∶3 anti-cyclin B (F2F4 monoclonal developed by P. O'Farrell, Developmental Studies Hybridoma Bank). All incubations were 2–3 hrs at room temperature except PLK-1 and cyclin B, which were incubated overnight at 4°C and room temperature, respectively.
Secondary antibodies from Invitrogen include goat anti-rabbit AlexaFluor 488-labeled IgG (used at 1∶100), goat anti-rat AlexaFluor 488-labeled IgG (used at 1∶100) and goat anti-mouse AlexaFluor 488-labeled IgG (used at 1∶100). Affinity purified secondary antibodies from Jackson Immunoresearch Laboratories include goat anti-rabbit TRITC-labeled IgG (used at 1∶100) and goat anti-mouse FITC-labeled IgG (used at 1∶100). DNA was visualized using the DNA dye DAPI at 0.1 µg/ml. Slides were prepared with either VectaShield (Vector Labs) or GelMount (Biomedia Corp.) as a combined mounting and anti-fade media.
Images were acquired via either confocal microscopy using a Leica TCSNT microscope, epifluorescence microscopy using a Zeiss Axiovert200M coupled with deconvolution via Slidebook 4.2 software (Intelligent Imaging Innovations), or DIC and epifluorescence on an Olympus BX60 microscope equipped with a Cooke Sensicam. Images acquired by confocal microscopy include those to visualize fibrillarin, HIM-10, SPE-11, HCP-4, and HCP-3. For deconvolution, images were acquired at 2×2 binning and 0.2 µm step sizes through each gonad and processed using either constrained iterative or nearest neighbors deconvolution. Images obtained via deconvolution include lamin, SYP-1, HIM-3, AIR-2, RNA pol II CTD(ser 2), SPD-2, and HCP-2. Epifluorescence images include pHisH3-ser10, α-tubulin and PLK-1.
Shakes D.C., Wu J.C., Sadler P.L., LaPrade K., Moore L.L., Noritake A, & Chu D.S. (2009). Spermatogenesis-Specific Features of the Meiotic Program in Caenorhabditis elegans. PLoS Genetics, 5(8), e1000611.
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Publication 2009

Most recents protocols related to «Fibrillarin»

1
Visualizing Chromosomes and Fibrillarin
Not available on PMC !
Flow sorted nuclei were mounted in polyacrylamide gel according to Nemecǩováet al. (2020) (link). To visualize 45S rDNA, chromosome 2 and chromosome 9, and fibrillarin, staining procedures and washes were performed according to Nemecǩovaé t al. ( 2020). Primary antibody anti-fibrillarin was diluted at 1:100 (ab4566, Abcam, Cambridge, UK). The hybridization mix for FISH contained 400 ng of individual probes.
Doležalová A., Beránková D., Koláčková V, & Hřibová E. (2024). Insight into chromatin compaction and spatial organization in rice interphase nuclei. Frontiers in plant science, 15.
Publication 2024
2
Immunofluorescent Quantification of Nucleolar and p53 Markers
CON and ALC fetuses, generated as above, were collected at E14.5 and their brains were fixed in 4% paraformaldehyde and preserved in 30% sucrose/PBS for cryoprotection. Heat-mediated antigen retrieval was performed on coronal sections (20 µm) with citrate antigen retrieval buffer (10 mM sodium citrate, 0.05% Tween-20, pH 6.0) using the steamer method. Fibrillarin-positive nucleoli were detected using rabbit anti-mouse fibrillarin (#ab5821, 1:1500; Abcam, Cambridge, UK) and visualized using Alexa-488 (#A-11094; Invitrogen; Waltham MA, USA)-labeled secondary antibody. P53-positive nuclei were detected using anti-rabbit p53 (1:200, R135, gift of N. Krupenko). Nuclei were visualized using 4′,6-diamidino-2-phenylindole (DAPI) (1:2000; Invitrogen) and mounted with Fluoromount-G (SouthernBiotech, Birmingham, AL, USA). The absence of the primary antibody served as a negative control. Digital images of equivalent cortical regions were captured under uniform exposure. The number and area of fibrillarin+ puncta per DAPI+ nucleus were quantified within the ventricular zone using CellProfiler 4.2.1 [35 (link)] (www.cellprofiler.org; accessed 24 September 2021). In brief, the number and area of DAPI+ nuclei and fibrillarin+ nucleoli were defined within 66 × 66 µm regions using the CellProfiler thresholding feature; the punctate nucleoli were further defined using the Enhance feature to remove background and the MaskImage feature to remove the overlapping DAPI signal. The number, area, and intensity of fibrillarin+ nucleoli were calculated for individual nuclei, counting 37–53 nuclei per cortex. The number of p53+ nuclei was quantified using ImageJ (v.54.1) [36 ] and sampling three sections per cortex. One fetus per litter was sampled from 4 CON and 5 ALC litters.
Huang Y., Flentke G.R., Rivera O.C., Saini N., Mooney S.M, & Smith S.M. (2024). Alcohol Exposure Induces Nucleolar Stress and Apoptosis in Mouse Neural Stem Cells and Late-Term Fetal Brain. Cells, 13(5), 440.
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Publication 2024
3
Quantitative Analysis of Nucleolar Proteins
Cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100, and blocked using 1% bovine serum albumin and 1% heat-inactivated goat serum in phosphate-buffered saline (PBS). Primary antibodies were directed against fibrillarin (#ab5821; 1:500), nucleolin (#ab22758, 1:250), UBF (#ab61205, 1:250; all from Abcam) or p53 (1:200, R135, gift of N. Krupenko), followed by secondary antibodies conjugated to Alexa-488 (1:1000, Invitrogen); nuclei were visualized using DAPI. Digital images were made under uniform exposure, capturing three images per sample, and the number of nucleoli per nucleus and percentage of fibrillarin+, nucleolin+, or UBF+ nucleolar area per DAPI+ nucleus were calculated using ImageJ [36 ].
Huang Y., Flentke G.R., Rivera O.C., Saini N., Mooney S.M, & Smith S.M. (2024). Alcohol Exposure Induces Nucleolar Stress and Apoptosis in Mouse Neural Stem Cells and Late-Term Fetal Brain. Cells, 13(5), 440.
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Publication 2024
4
Antibodies Used in Cell Studies
Antibodies used in this study include α-tubulin (immunofluorescent staining: AC007, ABclonal; Western blot: AC012, ABclonal), DDX21 (sc-376953, Santa Cruz Biotechnology), β-actin (AC026, ABclonal), Fibrillarin (A0850, ABclonal), GFP (HT801, TransGen Biotech), Cyclin B1 (sc-245, Santa Cruz Biotechnology), NOP58 (A4749, ABclonal) and Flag (Western blot: HT201, TransGen Biotech; immunofluorescent staining: F3165, Sigma-Aldrich). Thymidine (T1895), actinomycin d (SBR00013) and nocodazole (N219) were purchased from Sigma-Aldrich.
Jiang Y., Sun S., Liu X., Su K., Zhang C., Zhang P., Zhao Z., Su Y., Wang C, & Du X. (2024). U3 snoRNA inter-regulates with DDX21 in the perichromosomal region to control mitosis. Cell Death & Disease, 15(5), 342.
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Publication 2024
5
Silencing of DDX21, Fibrillarin, and U3 snoRNA
All siRNAs and ASOs were purchased from GenePharma (Shanghai, China). For silencing DDX21, siRNA-2 5’-CGGGAAUUAAGUUCAAACGAA-3’, siRNA-a 5’-GAGACUUAAUACUGAGCAAUG-3’ and siRNA-b 5’-GGGUUGUAAUACAGUUUAUAC-3’ were used. For silencing Fibrillarin, siRNA 5’- CGAGAGAUGUGUGUUGAUA-3’ was used. For the knockdown of U3 snoRNA, U3 ASO mUmUmCmGmGTGCTCTACACmGmUmUmCmA was used as described previously [53 (link)]. N and mN are deoxynucleotide and 2′-O-methoxyethylribonucleotide, respectively. Phosphodiester backbones are phosphorothioates.
Jiang Y., Sun S., Liu X., Su K., Zhang C., Zhang P., Zhao Z., Su Y., Wang C, & Du X. (2024). U3 snoRNA inter-regulates with DDX21 in the perichromosomal region to control mitosis. Cell Death & Disease, 15(5), 342.
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Publication 2024

Top products related to «Fibrillarin»

1
Ab4566 by Abcam
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Ab4566 is a primary antibody produced in rabbit. It is designed for use in immunohistochemistry applications.
2
Fibrillarin by Abcam
Sourced in United States
Fibrillarin is a highly conserved nucleolar protein that is involved in the processing and modification of ribosomal RNA (rRNA). It plays a crucial role in the biogenesis of ribosomes, which are the cellular machines responsible for protein synthesis.
3
Ab5821 by Abcam
Sourced in United Kingdom, United States
Ab5821 is a laboratory product made by Abcam. It serves a core function, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
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Anti fibrillarin by Abcam
Sourced in United States
Anti-fibrillarin is a laboratory reagent used for the detection and analysis of fibrillarin, a protein involved in the biogenesis of ribosomes in eukaryotic cells. It is typically used in immunoassays and other biochemical techniques to identify and quantify the presence of fibrillarin in various biological samples.
5
Fibrillarin by Cell Signaling Technology
Sourced in United States
Fibrillarin is a nuclear protein that functions as a methyltransferase, responsible for the methylation of pre-rRNA molecules. It plays a crucial role in the biogenesis of ribosomal subunits and the processing of small nucleolar RNAs (snoRNAs).
6
Fibrillarin by Santa Cruz Biotechnology
Sourced in United States
Fibrillarin is a nucleolar protein that is involved in the processing and methylation of ribosomal RNA (rRNA) during the early stages of ribosome biogenesis. It is a highly conserved protein found in eukaryotic cells and is essential for cell viability.
7
Anti flag by Merck Group
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Anti-FLAG is a lab equipment product used for the detection and purification of proteins tagged with the FLAG epitope. It functions as an affinity reagent that binds to the FLAG tag, enabling the isolation and identification of the tagged proteins.
8
Anti fibrillarin by Santa Cruz Biotechnology
Sourced in China, United States
Anti-Fibrillarin is a laboratory product offered by Santa Cruz Biotechnology. It is a protein-specific antibody that specifically recognizes and binds to the fibrillarin protein, which is a component of the small nucleolar ribonucleoprotein (snoRNP) complex. The core function of this product is to provide a tool for the detection and study of fibrillarin in various biological samples and applications.
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Vectashield by Vector Laboratories
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Vectashield is a non-hardening, aqueous-based mounting medium designed for use with fluorescent-labeled specimens. It is formulated to retard photobleaching of fluorescent dyes and provides excellent preservation of fluorescent signals.
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Alexa 488 by Thermo Fisher Scientific
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Alexa 488 is a fluorescent dye used in various biological applications. It is a brighly fluorescent, green-emitting dye with excitation and emission maxima at 495 and 519 nm, respectively. Alexa 488 can be conjugated to biomolecules such as proteins, antibodies, or nucleic acids to enable their detection and visualization.

More about "Fibrillarin"

Fibrillarin is a critical nucleolar protein that plays a central role in ribosome biogenesis and cellular function.
This highly conserved protein is involved in the processing and methylation of pre-ribosomal RNA, essential steps in the production of mature ribosomes.
Fibrillarin is found in eukaryotes and archaea, underscoring its fundamental importance in cellular machinery.
Disruptions to Fibrillarin and its associated pathways have been implicated in various disease states, including cancer and autoimmune disorders.
Understanding the intricacies of Fibrillarin is crucial for unraveling fundamental cellular processes and developing potential therapeutic interventions.
Researchers studying Fibrillarin can leverage Ab4566, a specific antibody that recognizes this target, as well as Ab5821, an Anti-Fibrillarin antibody.
These tools, along with the Anti-FLAG and Vectashield reagents, can be used to visualize and analyze Fibrillarin localization and expression patterns using techniques like immunofluorescence microscopy.
The Alexa 488 fluorescent label is also commonly employed to tag and detect Fibrillarin in cellular studies.
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This tool can help you easily locate the best protocols from literature, preprints, and patents, allowing you to discover the most reliable and effective methods to advance your Fibrillarin studies.
Leverage PubCompare.ai's powerful features to take your Fibrillarin research to new heights and contribute to the growing understanding of this crucial cellular component.