Anti-Nucleophosmin antibody [3A9F1]

• ICC

AbReview23395****

Immunocytochemistry - Anti-Nucleophosmin antibody [3A9F1] (AB86712)

ab86712 (1/2000) staining Nucleophosmin in assynchronous HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus. For further experimental details, please refer to abreview. (Please note that this particular image is a 3D projection of the data).

This image is part of an abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada

• ICC

Lab

Immunocytochemistry - Anti-Nucleophosmin antibody [3A9F1] (AB86712)

ab86712 stained HeLa cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab86712 at 5μg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43μM for 1hour at room temperature.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Nucleophosmin antibody [3A9F1] (AB86712)

Overlay histogram showing HeLa cells stained with ab86712 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab86712, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

• WB

Project8936****

Western blot - Anti-Nucleophosmin antibody [3A9F1] (AB86712)

All lanes:

Western blot - Anti-Nucleophosmin antibody [3A9F1] (ab86712) at 10 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

U2OS (Human osteosarcoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-hrp-preadsorbed-ab97040'>ab97040</a>) at 1/5000 dilution

Predicted band size: 32 kDa

Observed band size: 100 kDa,150 kDa,37 kDa,55 kDa

true

Exposure time: 1min