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 kDa. In ICC/IF, this antibody shows prominent specular nucleolar staining. Use in Immunohistochemistry whole mount reported in scientific literature (PMID 24722283)
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.
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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).
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FAQs for Fibrillarin (2)
In the immunohistochemistry paraffin section protocol....the PBS buffer....should it be at certain pH?
For IHC application, different labs cites the pH of PBS buffer in the range of 7.1 - 7.6 but most commonly used pH is pH 7.4 (this is what we use in our lab). You may use this PBS (pH 7.4) for making permeablization buffer, antibody diluent buffers and for making wash buffer. For more on the protocol that we use in our lab and for IHC-P troubleshooting suggestions, you may visit: IHC-P Protocol and IHC-P Troubleshooting.
I was wondering what does Format 7C mean as far as anitbodies go?
7C is a fluorophore - it is useful for FLOW. The details of the fluor are (A=425, E=500)