Products:Tags & Cell Markers >> Subcellular Markers >> Nucleus >> Nucleolus
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ab13745 |
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ab13745 |
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Read our guarantee »Publishing research using ab5821? Please let us know so that we can cite the reference in this datasheet
ab5821 has been referenced in 29 publications.
Publishing research using ab5821? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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.
Luke Hughes-Davies and Rhiannon Jade, Gurdon Institute, Cambridge, UK
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.
This image is courtesy of an Abreview submitted by Dr Tom Donndelinger
All lanes : Anti-Fibrillarin antibody - Nucleolar Marker (ab5821) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 :
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 34 kDa
Observed band size : 37 kDa (why is the actual band size different from the predicted?)
Additional bands at : 42 kDa,70 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 3 minutes
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