Fibrillarin Antibody (38F3)

Novus Biologicals | Catalog # NB300-269

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine, Bovine, C. elegans, Chicken, Drosophila, Equine, Plant, S. pombe, Yeast

Cited:

Human, Mouse, Insect - Drosophila, Nematode - Caenorhabditis elegans, Plant, S. pombe

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Immunohistochemistry Whole-Mount, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Knockdown Validated

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Immunohistochemistry Whole-Mount, Western Blot, Immunoblotting, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 38F3
View all Fibrillarin Primary Antibodies »

Product Specifications

Immunogen

Yeast nuclear preparation (S. cerevisiae).

Reactivity Notes

C. elegans and plant reactivity reported in scientific literature (PMID: 24722283 and 27588463 respectively). Use in S. pombe reported in scientific literature (PMID:31855181).

Localization

Nucleus > nucleolus. Note: Fibrillar region of the nucleolus.

Marker

Nucleolar Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Theoretical MW

34.5 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for Fibrillarin Antibody (38F3)

Immunocytochemistry/ Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269]

Immunocytochemistry/ Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269]

Immunocytochemistry/Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] - High magnification confocal image of HeLa cells stained with fibrillarin antibody, dilution 1:100 from concentrated tissue culture media (Green), and costained with chicken vimentin pAb (Red), dilution 1:1,000. Nuclear DNA is revealed with the DAPI stain (Blue). The fibrillarin antibody shows strong staining of nucleoli in the nucleus, while the vimentin antibody reveals cytoplasmic intermediate filaments.
Western Blot: Fibrillarin Antibody (38F3) [NB300-269]

Western Blot: Fibrillarin Antibody (38F3) [NB300-269]

Western Blot: Fibrillarin Antibody (38F3) [NB300-269] - Analysis of lysates of cell fractions probed with Fibrillarin antibody, dilution 1:500 (Green): [1] protein standard, [2] C6 cytosol, [3] C6 nuclear, [4] HEK293 cytosol, [5] HEK293 nuclear, [6] NIH-3T3 cytosol and [7] NIH-3T3 nuclear fractions. The band at 37kDa corresponds to the fibrillarin protein detected exclusively in the nuclear fractions.
Immunocytochemistry/ Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269]

Immunocytochemistry/ Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269]

Fibrillarin-Antibody-38F3-Immunocytochemistry-Immunofluorescence-NB300-269-img0012.jpg
Immunohistochemistry-Paraffin: Fibrillarin Antibody (38F3) [NB300-269]

Immunohistochemistry-Paraffin: Fibrillarin Antibody (38F3) [NB300-269]

Immunohistochemistry-Paraffin: Fibrillarin Antibody (38F3) [NB300-269] - IHC staining of Fibrillarin in human ovarian cancer using DAB with hematoxylin counterstain.
Immunocytochemistry/ Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269]

Immunocytochemistry/ Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269]

Immunocytochemistry/Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] - Mouse embryonic fibroblast fibrillarin stained with fibrillarin antibody and Alexa-594 secondary antibody. ICC/IF image submitted by a verified customer review.
Flow Cytometry: Fibrillarin Antibody (38F3) [NB300-269]

Flow Cytometry: Fibrillarin Antibody (38F3) [NB300-269]

Flow Cytometry: Fibrillarin Antibody (38F3) [NB300-269] - Fibrillarin antibody was tested at 1:400 in HEK293 cells using an Alexa Fluor 488 secondary (shown in green) alongside unstained cells (shown in red).
Western Blot: Fibrillarin Antibody (38F3) [NB300-269]

Western Blot: Fibrillarin Antibody (38F3) [NB300-269]

Western Blot: Fibrillarin Antibody (38F3) [NB300-269] - Western blot analysis of Fibrillarin expression on yeast protein extracts using NB300-269.
Immunocytochemistry/ Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269]

Immunocytochemistry/ Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269]

Immunocytochemistry/Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] - Fibrillarin immunofluorescence in fibroblasts. ICC/IF image submitted by a verified customer review.
Immunocytochemistry/ Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269]

Immunocytochemistry/ Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269]

Immunocytochemistry/Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] - IF Confocal analysis of MCF7 cells using Fibrillarin antibody (NB300-269, 1:5). An Alexa Fluor 488-conjugated Goat to mouse IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).
Fibrillarin Antibody (38F3)

Immunocytochemistry/ Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] -

Immunocytochemistry/ Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] - The nuclei of hth mutants have dispersed rDNA foci & ectopic nucleoli & produce more rRNA.A) Wild type nuclei stained with anti-Fibrillarin (red) & a specific probe that recognizes the rDNA genomic region (green). B) hth mutant nuclei stained as in A. The mutant nuclei exhibit an increased number of nucleoli with different sizes. C) Quantification of nuclear dots marked with both the rDNA probe & anti-Fibrillarin in wild type & mutant nuclei. 45 nuclei were quantified for each genotype. (p-value: 0,00010132, calculated using T-Test). D) qRT-PCR comparing the levels of 18S RNA transcripts in wild type & Dfhth mutant embryos. The experiment was performed three independent times to be significant (p-value: 0,01046, calculated using T-Test). Error bars in the graph show the standard deviation & have being calculated as in Fig. 1E. The experiment was normalized using the bicoid (bcd) mRNA. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25794008), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Fibrillarin Antibody (38F3)

Immunocytochemistry/ Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] -

Immunocytochemistry/ Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] - The nuclei of hth mutants have dispersed rDNA foci & ectopic nucleoli & produce more rRNA.A) Wild type nuclei stained with anti-Fibrillarin (red) & a specific probe that recognizes the rDNA genomic region (green). B) hth mutant nuclei stained as in A. The mutant nuclei exhibit an increased number of nucleoli with different sizes. C) Quantification of nuclear dots marked with both the rDNA probe & anti-Fibrillarin in wild type & mutant nuclei. 45 nuclei were quantified for each genotype. (p-value: 0,00010132, calculated using T-Test). D) qRT-PCR comparing the levels of 18S RNA transcripts in wild type & Dfhth mutant embryos. The experiment was performed three independent times to be significant (p-value: 0,01046, calculated using T-Test). Error bars in the graph show the standard deviation & have being calculated as in Fig. 1E. The experiment was normalized using the bicoid (bcd) mRNA. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25794008), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Fibrillarin Antibody (38F3)

Immunocytochemistry/ Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] -

Immunocytochemistry/ Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] - The nuclei of hth mutants have dispersed rDNA foci & ectopic nucleoli & produce more rRNA.A) Wild type nuclei stained with anti-Fibrillarin (red) & a specific probe that recognizes the rDNA genomic region (green). B) hth mutant nuclei stained as in A. The mutant nuclei exhibit an increased number of nucleoli with different sizes. C) Quantification of nuclear dots marked with both the rDNA probe & anti-Fibrillarin in wild type & mutant nuclei. 45 nuclei were quantified for each genotype. (p-value: 0,00010132, calculated using T-Test). D) qRT-PCR comparing the levels of 18S RNA transcripts in wild type & Dfhth mutant embryos. The experiment was performed three independent times to be significant (p-value: 0,01046, calculated using T-Test). Error bars in the graph show the standard deviation & have being calculated as in Fig. 1E. The experiment was normalized using the bicoid (bcd) mRNA. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25794008), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Fibrillarin Antibody (38F3)

Western Blot: Fibrillarin Antibody (38F3) [NB300-269] -

In vitro VM triggers nuclear YAP1 expression in U87 and U251 glioblastoma cells and is inhibited by Hippo pathway pharmacological targeting. (A) Human U87 and U251 glioblastoma cells were seeded either as monolayers (2D) or on top of Cultrex (3D) to generate capillary‐like structures. Subcellular fractionation was performed to isolate the cytosol and nuclear fractions from 2D or 3D cells. Representative blots for YAP1, Fibrillarin and GAPDH are presented from three independent fractionations. (B) Cytosolic (Cyt) and nuclear (Nuc) fractions were isolated from U87 glioblastoma cells seeded on Cultrex and treated with either vehicle or 10 μM of the indicated Hippo pathway inhibitors for 24 h. Representative blots for YAP1 and Fibrillarin are presented from two independent fractionations. (C) Total RNA was extracted from U87 cells cultured as in (A). RT‐qPCR was performed and relative gene expression of CTGF, Cyr61 and PPIA normalised over GAPDH. Data presented are representative triplicates from two independent experiments (*p < 0.05). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39718433), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Fibrillarin Antibody (38F3)

Western Blot: Fibrillarin Antibody (38F3) [NB300-269] -

In vitro VM triggers nuclear YAP1 expression in U87 and U251 glioblastoma cells and is inhibited by Hippo pathway pharmacological targeting. (A) Human U87 and U251 glioblastoma cells were seeded either as monolayers (2D) or on top of Cultrex (3D) to generate capillary‐like structures. Subcellular fractionation was performed to isolate the cytosol and nuclear fractions from 2D or 3D cells. Representative blots for YAP1, Fibrillarin and GAPDH are presented from three independent fractionations. (B) Cytosolic (Cyt) and nuclear (Nuc) fractions were isolated from U87 glioblastoma cells seeded on Cultrex and treated with either vehicle or 10 μM of the indicated Hippo pathway inhibitors for 24 h. Representative blots for YAP1 and Fibrillarin are presented from two independent fractionations. (C) Total RNA was extracted from U87 cells cultured as in (A). RT‐qPCR was performed and relative gene expression of CTGF, Cyr61 and PPIA normalised over GAPDH. Data presented are representative triplicates from two independent experiments (*p < 0.05). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39718433), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for Fibrillarin Antibody (38F3)

Application
Recommended Usage

Flow Cytometry

1:20-1:400

Immunocytochemistry/ Immunofluorescence

1:500-1:1000

Immunohistochemistry

1:500-1:1000

Immunohistochemistry-Frozen

1:500-1:1000

Immunohistochemistry-Paraffin

1:500-1:1000

Western Blot

1:1000-1:5000
Application Notes
This Fibrillarin Antibody (38F3) is useful for Western blot, Immunocytochemistry/Immunofluorescence, Flow Cytometry and Immunohistochemistry on both paraffin-embedded and frozen sections. In WB a band can be seen at approximately 34.5 kDa. In ICC/IF, this antibody shows prominent specular nucleolar staining.

Use in IHC-WhMt and IHC-Fr reported in scientific literature (PMID: 24722283 and 21539824 respectively). Use in Immunohistochemistry-paraffin reported in scientific literature (PMID: 23542174).

Reviewed Applications

Read 4 reviews rated 5 using NB300-269 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Unpurified

Formulation

Supplied as concentrated hybridoma cell culture media.

Preservative

0.035% Sodium Azide

Concentration

This product is unpurified. The exact concentration of antibody is not quantifiable.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Fibrillarin

Fibrillarin / Nop1p, was originally identified as a nucleolar protein of baker's yeast, Saccharomyces cerevisiae (accession P15646). The Nop1p protein is essential for yeast viability and is localized in the nucleoli. The human homologue of Nop1p is Fibrillarin (accession P22087), a component of the nucleolar small nuclear ribonucleoprotein (snRNP) particle. The human Fibrillarin gene is located on chromosome 19 (19q13.1). Fibrillarin proteins have been cloned and sequenced from several other species (mouse, accession P35550, Xenopus accession P22232, C. elegans accession Q22053, and S. pombe accession P35551). The N terminal 80 amino acids contain multiple copies based on the peptide RGG, and the remaining 240 amino acids consist of the Fibrillarin domain. A Fibrillarin homologue has also been identified in the genome of the archean Methanococcus (accession NC_000909). This protein lacks the RGG rich N-terminal extension but is clearly homologous to the other sequences throughout the entire Fibrillarin domain. The structure of this molecule has been determined and shown to consist of 2 extended beta-sheets flanked by 4 alpha-helixes. Patients with the autoimmune disease scleroderma often have strong circulating autoantibodies to a 34kDa protein which was subsequently found to be Fibrillarin. Fibrillarin is an excellent marker for the nucleolus.

Alternate Names

EC 2.1.1,34-kD nucleolar scleroderma antigen, EC 2.1.1.-, EC 2.1.1.37, FIB, FIB1,34 kDa nucleolar scleroderma antigen, fibrillarin, FLRNrRNA 2'-O-methyltransferase fibrillarin, RNA, U3 small nucleolar interacting protein 1, RNU3IP1

Entrez Gene IDs

2091 (Human); 14113 (Mouse); 292747 (Rat)

Gene Symbol

FBL

Additional Fibrillarin Products

Product Documents for Fibrillarin Antibody (38F3)

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for Fibrillarin Antibody (38F3)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for Fibrillarin Antibody (38F3)

Customer Reviews for Fibrillarin Antibody (38F3) (4)

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Customer Images

Showing  1 - 4 of 4 reviews Showing All
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  • Fibrillarin Antibody (38F3)
    Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: Mouse embryonic fibroblasts and fibroblasts
    Species: Mouse
    Verified Customer | Posted 06/14/2021
    immunofluorescence of fibrillarin in mouse embryonic fibroblast using Alexa-594 secondary antibody
    Fibrillarin Antibody (38F3) NB300-269
  • Fibrillarin Antibody (38F3)
    Name: Varnesh Tiku
    Application: Western Blot
    Sample Tested: Tissue Lysates and 4% PFA-PBS fixed frozen sections from murine brain
    Species: C. elegans
    Verified Customer | Posted 01/09/2019
  • Name: Bryan Tinsley
    Application: Immunocytochemistry
    Sample Tested:
    Species: Human
    Verified Customer | Posted 06/09/2014
    IF Confocal analysis of MCF7 cells using Fibrillarin antibody (NB300-269, 1:5).
    Fibrillarin Antibody (38F3) NB300-269
  • Fibrillarin Antibody (38F3)
    Name: Anonymous
    Application: Western Blot
    Sample Tested: fibroblasts
    Species: Other
    Verified Customer | Posted 04/24/2014
    Fibrillarin immunofluorescence in fibroblasts

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Protocols

View specific protocols for Fibrillarin Antibody (38F3) (NB300-269):


Materials

1) 1 Phosphate buffered saline (pH 7.6): NaCl 137mmol/L, KCl 2.7mmol/L, Na2HPO4 4.3mmol/L, KH2PO4 1.4 mmol/L
2) Citrate buffer, 0.01 M, pH6.0, Sodium Citrate 3g, Citric acid 0.4g
3) 3% Hydrogen peroxide
4) Primary antibody
5) Blocking serum (normal serum)
6) Biotinylated secondary antibody
7) DAB staining kit

Methods

1. Dewax and hydration of slides using xylene and EtOH:
Dry slides for 20 min in a 60 C oven
Add Xylene, 2 x 10 min
100%, 95%, 80%, and 70% EtOH, 5 min each EtOH concentration
Rinse in PBS, 5'

2 Antigen retrieval method (only for paraffin slides)
1a. High-pressure antigen retrieval procedure (recommended method)
Place slides in a glass slide holder (ensure that the slide holder is completely filled with slides, slides without sections if necessary, to ensure even heating. The entire slide holder is immersed in 1000 ml of Citrate buffer (0.01M, pH6.0) within a pressure cooker
Once steam is produced, and ONLY when steam is visible, from the pressure cooker (usually 15-20 min), the required high-pressure will have been reached, and slides will be incubated for 2 min.
Turn off heat, and allow buffer and slides to cool to room temperature
Slides are then rinsed in PBS for 5 minutes
2. Add 3% hydrogen peroxide solution, 10'at RT, then PBS, 3X5'
3. Normal blocking serum, 20'at RT
4. Incubate with Primary Ab, 4C overnight or 1.5 hours at 37C
5. Rinse with PBS, 3 X 5' each rinse
6. Add Biotin-conjugated second antibody, 10'at RT
7. Rinse with PBS, 3 X 5' each rinse
8. Add Streptavidin-Peroxidase, 10'at RT
9. Rinse with PBS, 3 X 5' each rinse
10. Staining with DAB solution, 2-5'under microscope
11. Stop the reaction by washing in tap water
12. Counterstain in Haematoxylin for 3-5 minutes
13. 75%, 80%, 95% and 100% ethanol, 5x2', xylene 2 x 10'

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FAQs

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