EnCor Biotechnology Inc.

Rabbit Polyclonal Antibody to Fibrillarin Cat# RPCA-Fib

$300.00
Description

      The RPCA-Fib antibody was made against recombinant human fibrillarin expressed in and purified from E. coli and is superior on western blots of mammalian samples to the widely used MCA-38F3 antibody, which was originally raised against yeast Nop1p and later found to recognize fibrillarin, the mammalian homologue of the yeast protein. However MCA-38F3 has been documented to be usable as a marker of nucleoli in a wide variety of species which has not so far been documented with this reagent. The chicken antibody works well for IF and ICC but is not recommended for IHC. We have also produced a mouse monoclonal MCA-4A4, both made against recombinant human fibrillarin. Mouse select image above left for larger view.
Amount: 100µL of 1mg/mL
Amount: 100µL of 1mg/mL
Immunofluorescent analysis of HeLa cells stained with rabbit pAb to fibrillarin, RPCA-Fib, dilution 1:5,000 in red, and chicken pAb to vimentin, CPCA-Vim, dilution 1:10,000 in green. The first image was taken at high magnification using a 100X objective, the second is a lower power overview taken with a 40X lens. The blue is DAPI staining of nuclear DNA. The RPCA-Fib antibody detects protein expressed in nucleoli of cells, while the CPCA-Vim antibody produces strong staining of cytoplasmic intermediate filaments.
Western blot analysis of cell lysates using rabbit pAb to fibrillarin RPCA-Fib, dilution 1:5,000, in green: lane [1] protein standard (red), and extracts of [2] NIH/3T3, [3] HEK293, [4] HeLa, [5] SH-SY5Y, [6] C6, and [7] COS-1 cells . The strong band at ~35 kDa corresponds to the fibrillarin protein.

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Name: Fibrillarin, chicken polyclonal antibody, Cat# CPCA-Fib
Immunogen: Full length human fibrillarin expressed in and purified from E. coli.
HGNC Name: FBL
UniProt: P22087
Molecular Weight: 34.5kDa
Host: Chicken
Species Cross-Reactivity: Human, Rat, Mouse, Dog, Horse
RRID: AB_2572216
Format: Purified antibody at 1mg/mL in 50% PBS, 50% glycerol plus 5mM sodium azide
Applications: WB, IF/ICC
Recommended Dilutions: WB: 1:2,000-1:5,000. IF/ICC: 1:2,000-1:5,000. IHC not recommended
Storage: Store at 4°C

      Fibrillarin is a highly conserved component of a nucleolar small ribonucleoprotein complex in mammals, involved in the processing of ribosomal RNA during ribosomal biogenesis. The protein runs at ~35kDa on SDS-PAGE and is very rich in basic amino acids having a PI of 9.8. Fibrillarin was originally identified in humans since autoantibodies staining nucleoli were seen in some patients with the autoimmune disease scleroderma (1). Subsequently the protein fibrillarin was found to be the human homologue of Nop1p, a Saccharomyces cerevisiae nucleolar protein, the two proteins being 67% identical (2,3). We have generated an alignment of the sequences of fibrillarin and homologues downloadable from here. The fibrillarin molecule consists of an N-terminal glycine and arginine rich region followed by a highly conserved globular domain. Embryonic knockout of the fibrillarin gene in mice is lethal, suggesting fundamental importance of this protein (4). Autoantibodies to fibrillarin are also seen in patients with the autoimmune disease systemic sclerosis (5).

      This antibody has been tested on formalin fixed and paraffin embedded samples for IHC, and is not recommended for this purpose.

1. Aris JP and Blobel G. Identification and characterization of a yeast nucleolar protein that is similar to a rat liver nucleolar protein. J. Cell Biol. 107:17-31 (1988).
2. Aris JP and Blobel G. cDNA cloning and sequencing of human fibrillarin, a conserved nucleolar protein recognized by autoimmune antisera. Proc. Natl. Acad. Sci. 88:931-5 (1991).
3. Ochs RL, Lischwe MA, Spohn WH, Busch H. Fibrillarin: a new protein of the nucleolus identified by autoimmune sera. Biol. Cell. 54:123-33 (1985).
4. Newton K, Petfalski E, Tollervey D, Caceres JF. Fibrillarin is essential for early development and required for accumulation of an intron-encoded small nucleolar RNA in the mouse. Mol. Cell Biol. 23:8519-27 (2003).
5. Okano Y, Steen VD, Medsger TA. Autoantibody to U3 nucleolar ribonucleoprotein (fibrillarin) in patients with systemic sclerosis. Arth. Rheum. 35:95-100 (1992).

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