||Mouse Anti-Nop1p/Fibrillarin mAb; Anti Nop1p/Fibrillarin
||Nop1p/Fibrillarin was originally identified as a nucleolar protein of bakers 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 homologues to the other sequences throughout all of the fibrillarin domain. The structure of this molecule has been determined and shown to consist of 2 extended b-sheets flanked by 4 a-helixes. Patients with the autoimmune disease scleroderma often have strong circulating autoantibodies to a 34,000 Da protein which was subsequently found to be fibrillarin. Fibrillarin is an excellent marker for the nucleolus.
||Yeast fibrillarin, a highly conserved nucleolar protein present in small nucleolar ribonucleoprotein (snoRNP) complexes.
||1 mg/ml in PBS, pH 7.4 with 0.05% sodium azide. Frozen liquid
||Cross reacts with fibrillarin in human, rat, plant, Drosophia, C. elegans and S. pombe.
||Stable for 1 year at -20°C from date of shipment. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Aliquot to avoid repeated freezing and thawing. Aliquot will be stable at 4°C for 3 months.
||Briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container's cap because small volumes of antibody will occasionally become entrapped in the seal of the product vial during shipment and storage.
|Western blots of yeast protein samples: 1:1,000 (cell lysates) to 1:5,000 (nuclear fractions), followed by chemiluminescent detection (ECL).
Other (non-ECL) Western detection methods: 1:500 to 1:2,500
Detect mammalian fibrillarin on western blots by ECL: 1:250
Immunofluoscence on yeast cells: 1:500 to 1:2,500
IF of mammalian cells: 1:250.
Levitskii SA, et al. Identification of signal sequences determining the specific nucleolar localization of fibrillarin in HeLa cells. Mol. Biol. (Mosk). May-Jun 2004; 38(3): 483-492.
Newton K, et al. Fibrillarin is essential for early development and required for accumulation of an intron-encoded small nucleolar RNA in the mouse. Mol. Cell Biol. Dec 2003; 23(23): 8519-8527.