Anti-NPM1-mutated antibody [EPR30901-1]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-NPM1-mutated antibody [EPR30901-1] (AB326567)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized
293T cells transfected with empty vector containing a myc-HA-tag®
293T cells transfected with a human NPM1(mutation) expression vector containing a myc-HA-tag®
293T cells transfected with a human wild-type NPM1 expression vector containing a myc-HA-tag®
labelling NPM1-mutated with ab326567 at 1/500 (0.966 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing positive staining in 293T cells transfected with a human NPM1(mutation) expression vector containing a myc-HA-tag® (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-NPM1-mutated antibody [EPR30901-1] (AB326567)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized
293T cells transfected with a human NPM (mutation) expression vector containing a HA-Myc-tag® (Upper)
293T cells transfected with a human NPM (WT) expression vector containing a HA-Myc-tag® (Lower left)
293T cells transfected with an empty expression vector containing a myc-His-tag® (Lower right)
labelling NPM1-mutated with ab326567 at 1/5000 dilution (0.01ug) / Upper right and Lower left/right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• WB

Lab

Western blot - Anti-NPM1-mutated antibody [EPR30901-1] (AB326567)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab236632) (1 : 200000) (36KDa);Anti-HA tag® antibody- (ab236632) (1 : 1000)(30-100KDa).

All lanes:

Western blot - Anti-NPM1-mutated antibody [EPR30901-1] (ab326567) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a Myc-HA-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human wild-type NPM1 expression vector containing a Myc-HA-tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human NPM1(mutation) expression vector containing a Myc-HA-tag, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 30-100 kDa,36 kDa

false

Exposure time: 3s

• Dot

Lab

Dot Blot - Anti-NPM1-mutated antibody [EPR30901-1] (AB326567)

Dot blot analysis of NPM1-mutated using ab326567 at 1 : 1000 (0.483 μg/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane 1 : Human NPM1 mutant peptide

Lane 2 : Human NPM1 wide-type peptide

Lane 3 : Human NPM1 mutant C-terminus sequence

Exposure time : 10 seconds.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Dot Blot - Anti-NPM1-mutated antibody [EPR30901-1] (ab326567) at 1/1000 dilution

Lane 1:

Human NPM1 mutant peptide

Lane 2:

Human NPM1 wide-type peptide

Lane 3:

Human NPM1 mutant C-terminus sequence

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

Exposure time: 10s