Anti-Nup153 antibody [QE5] (ab24700)

ab24700 staining Nup153 in HeLa cells. The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab24700 at 0.4 µg/ml concentration and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at +4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (ab150117) at 2 µg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 2 µg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

ab24700 staining Nup153 in NIH3T3 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab24700 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 100% methanol (5 min).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

Immunocytochemical analysis of U2OS cells, labeling Nup153 with ab24700. Cells were formaldehyde fixed, permeabilized with NP40, and blocked with 3% BSA for 1 hour at 21°C. Immunostaining with ab24700 diluted 1/100 for 12 hours at 4°C.

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Human and mouse cells treated with different concentrations of ab24700. The cells were fixed in 4% formaldehyde and permeabilized in 0.2% Triton x100 for 10 minutes at room temperature, then washed 3 times in PBS. The fixation and permeabilization was carried out in a single step. The image shows clear staining of the nuclear envalope (green). The DNA is stained with DAPI (blue).

A: HeLa cells, ab24700 used at 4µg/ml
B: 3T3 cells, ab24700 used at 2µg/ml
C: 3T3 cells, ab24700 used at 4µg/ml
D: 3T3 cells, ab24700 used at 6.6µg/ml

Human and mouse cells treated with different concentrations of ab24700. The cells were fixed in 100% methanol for 20 minutes at -20°C, then washed once in PBS. The image shows clear staining of the nuclear envalope (green). The DNA is stained with DAPI (blue).

A: HeLa cells, ab24700 used at 4µg/ml
B: 3T3 cells, ab24700 used at 2µg/ml
C: 3T3 cells, ab24700 used at 4µg/ml
D: 3T3 cells, ab24700 used at 6.6µg/ml