Anti-Fibrillarin antibody - Nucleolar Marker

• Supplier: Abcam
• Catalog: ab5821
• Prices: $401.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: IHC ICC/IF ICC/IF IP IHC-P WB ICC/IF
• Species Reactivities: Mouse, Human, Xenopus, Drosophila
• Antigen: Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Fibrillarin
• Blocking Peptide: Human Fibrillarin peptide
• Description: Rabbit Polyclonal
• Gene: Fib
• Conjugate: Unconjugated

Product images

The cells were 100% methanol fixed (5 min) and thenincubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab37266, 1µg/ml, red) and (ab5821, 1µg/ml, blue) overnight at +4°C. The secondary antibodies were ab150115 Alexa Fluor® 647 goat anti-mouse IgG (H+L) used at 2µg/ml for 1h and ab175652 Alexa Fluor® 405 goat anti-rabbit IgG (H+L) used at 2µg/ml for 1h. 

ICC/IF image of ab5821 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab5821, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

Immunofluorescent imaging of human cells (U2OS) with ab5821 reveals highly specific localisation to the dense fibrillar component (DFC) of the nucleolus associated with the initial ribosomal RNA (rRNA) precursor. The nucleolar protein fibrillarin is located primarily in the DFC.  Blue is hoechst staining of the nucleus, green is ab5821 used at 1/100, merge image demonstrates exclusively nuclear localisation.   IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour.  Primary antibody used at 1/100 in 5% milk / 0.2% TWEEN for one hour, secondary antibody Alexa 488 for 30 minutes.  All blocking and incubation steps carried out at 37 degrees C.

ab5821 staining human liver tissue sections by IHC-P.  Sections were formaldehyde fixed, permeabilized in Triton-X and subjected to heat mediated antigen retrieval prior to blocking with 6% BSA for 2 hours at 23°C. The primary antibody was diluted 1/100 and incubated with the sample for 16 hours at 5°C.  ab6717 was used as the secondary antibody.See Abreview

All lanes : Anti-Fibrillarin antibody - Nucleolar Marker (ab5821) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 3 : Schneider L2 (Fruit fly (Drosophila melanogaster)) Whole Cell Lysate (ab14893)Lysates/proteins at 20 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilutionPerformed under reducing conditions.

Immunohistochemical analysis of salivary glands from Drosophila melanogaster larvae, staining Fibrillarin with ab5821. Tissues were fixed in 4% paraformaldehyde/PBS and 0.3% Triton-X/PBS before incubation with primary antibody.

Product References

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Tessarz P, Santos-Rosa H, Robson SC, Sylvestersen KB, Nelson CJ, Nielsen ML, Kouzarides T. Nature. 2014 Jan 23;505(7484):564-8.

Drosophila Spag is the homolog of RNA polymerase II-associated protein 3 (RPAP3) - Drosophila Spag is the homolog of RNA polymerase II-associated protein 3 (RPAP3)

Benbahouche Nel H, Iliopoulos I, Torok I, Marhold J, Henri J, Kajava AV, Farkas R, Kempf T, Schnolzer M, Meyer P, Kiss I, Bertrand E, Mechler BM, Pradet-Balade B. J Biol Chem. 2014 Feb 28;289(9):6236-47.

The Hmr and Lhr hybrid incompatibility genes suppress a broad range of - The Hmr and Lhr hybrid incompatibility genes suppress a broad range of

Satyaki PR, Cuykendall TN, Wei KH, Brideau NJ, Kwak H, Aruna S, Ferree PM, Ji S, Barbash DA. PLoS Genet. 2014 Mar 20;10(3):e1004240.

Nucleolar organization, ribosomal DNA array stability, and acrocentric chromosome - Nucleolar organization, ribosomal DNA array stability, and acrocentric chromosome

Stimpson KM, Sullivan LL, Kuo ME, Sullivan BA. PLoS One. 2014 Mar 24;9(3):e92432.

Fam118B, a newly identified component of Cajal bodies, is required for Cajal body - Fam118B, a newly identified component of Cajal bodies, is required for Cajal body

Li Y, Fong KW, Tang M, Han X, Gong Z, Ma W, Hebert M, Songyang Z, Chen J. J Cell Sci. 2014 May 1;127(Pt 9):2029-39.

Nucleolar integrity is required for the maintenance of long-term synaptic - Nucleolar integrity is required for the maintenance of long-term synaptic

Allen KD, Gourov AV, Harte C, Gao P, Lee C, Sylvain D, Splett JM, Oxberry WC, van de Nes PS, Troy-Regier MJ, Wolk J, Alarcon JM, Hernandez AI. PLoS One. 2014 Aug 4;9(8):e104364.

A separable domain of the p150 subunit of human chromatin assembly factor-1 - A separable domain of the p150 subunit of human chromatin assembly factor-1

Smith CL, Matheson TD, Trombly DJ, Sun X, Campeau E, Han X, Yates JR 3rd, Kaufman PD. Mol Biol Cell. 2014 Sep 15;25(18):2866-81.

RNA-Seq of the nucleolus reveals abundant SNORD44-derived small RNAs. - RNA-Seq of the nucleolus reveals abundant SNORD44-derived small RNAs.

Bai B, Yegnasubramanian S, Wheelan SJ, Laiho M. PLoS One. 2014 Sep 9;9(9):e107519.

Ultrastructure of cytoplasmic and nuclear inosine-5'-monophosphate dehydrogenase - Ultrastructure of cytoplasmic and nuclear inosine-5'-monophosphate dehydrogenase

Juda P, Smigova J, Kovacik L, Bartova E, Raska I. J Histochem Cytochem. 2014 Oct;62(10):739-50.

Plasmodium berghei MAPK1 displays differential and dynamic subcellular - Plasmodium berghei MAPK1 displays differential and dynamic subcellular

Wierk JK, Langbehn A, Kamper M, Richter S, Burda PC, Heussler VT, Deschermeier C. PLoS One. 2013;8(3):e59755.